Sample vessels

ABSTRACT

A device for processing a biological sample includes a processing unit having at least one opening to receive a sample vessel and a plurality of processing stations positioned along the opening. The processing stations each have a compression member adapted to compress the sample vessel within the opening and thereby move a substance within the sample vessel among the processing stations. An energy transfer element can be coupled to one or more of the processing stations for transferring thermal energy to the content at a processing station.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.10/241,816, filed Sep. 11, 2001, which is a continuation-in-part of U.S.patent application Ser. No. 09/782,732, filed Feb. 13, 2001, now U.S.Pat. No. 6,780,617, which claims the benefit of U.S. Provisional PatentApplication Ser. No. 60/259,025, filed Dec. 29, 2000. application Ser.No. 10/241,816 is also a continuation-in-part of U.S. patent applicationSer. No. 09/910,233, filed Jul. 20, 2001, now U.S. Pat. No. 6,748,332,which is a continuation of U.S. patent application Ser. No. 09/339,056,filed Jun. 23, 1999, now U.S. Pat. No. 6,318,191, which claims thebenefit of U.S. Provisional Patent Application Ser. No. 60/090,471,filed Jun. 24, 1998. application Ser. No. 10/241,816 further claims thebenefit of U.S. Provisional Patent Application Ser. No. 60/318,768,filed Sep. 11, 2001. Each of the aforementioned patent applications andpatents is incorporated herein by reference.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

Portions of the disclosed subject matter were made with governmentsupport under grant numbers 1R43HL65768, 1R43HL65867 and 1R43HL67568awarded by the National Institutes of Health. The government has certainrights in those portions.

BACKGROUND

As result of the Human Genome Project and other genetic research, atremendous amount of genomic and biomarker information is presentlyavailable to healthcare providers. Using molecular diagnostic testing,genomic and biomarker information can provide a resource to healthcareproviders to assist in the rapid and accurate diagnosis of illness.However, the development of diagnostic testing systems allowing the useof such genetic information, particularly in the clinical setting, hasfailed to match pace with the genetic research providing theinformation. Current diagnostic testing systems are mainly limited tolarge medical testing centers or research labs due to the high costsassociated with acquiring and operating the systems and the complexityof the molecular diagnostic assays being employed. These current systemsrequire a large initial capital investment and incur high costs forreagents, disposables, operation, maintenance, service and training.

Sample preparation and handling generally includes sample collection andany preprocessing required for subsequent biological and chemicalassays. Sample collection and handling is an important part of in vitrodiagnostic (IVD) testing, and is an important factor in determining thefeasibility of test automation. With the advancement of medicine, thenumber of possible assays available to perform is continuallyincreasing. In parallel, sample collection methods have evolved over thelast several decades. In the case of blood sample collection, forexample, disposable plastic syringes first replaced glass syringes toimprove safety. Later developments had vacuum tubes replacing thetraditional syringes to simplify the blood collection process. However,a vacuum tube is generally not suitable for use as an IVD test reactionchamber. Thus, a re-sampling process is necessary for delivery of thesample to distinct assay containers for each of a variety of IVD tests.Automation of these processes is a daunting task. Indeed, in largeclinical testing centers giant automation testing systems costingseveral million dollars are currently used. The major automated task inthese machines is liquid handling, which entails the pipetting of thesample from sample tubes to 96-well plates, the addition of thereagent(s) to the wells, as well as moving reaction mixtures from wellto well.

Recently, nanotechnology has emerged to revolutionize automation andtesting formats. In this direction, by using silicone micro-fabricationand etching technology, the lab-on-a-chip platform was developed in anattempt to integrate and miniaturize certain parts of the automationprocess into a chip with dimensions less than 2 mm by 2 mm. Liquidprocessing rates for certain lab-on-a-chip platforms can be on the scaleof nanoliters per second. However, it is often difficult for users tointerface with this type of platform to, for example, deliver the sampleto the chip.

Another concern of current sample handling devices is the large samplevolume routinely drawn from a patient for IVD testing. In the case ofblood sample collection, for example, a small vacuum tube may take closeto 5 ml whole blood. When multiple samples are required in the testingof various assays, several tubes of blood are frequently ordered.However, only a small amount is needed for each assay. The drawing of alarge volume of blood for multiple tests is a concern for pediatricpatients as it can lead to iron deficiency anemia. It is even morecritical for patients with pre-existing anemia or a bleeding disorder.

SUMMARY

The present invention provides sample processing devices and methodsthat facilitate the rapid analysis of biological samples, such as blood,saliva, or urine, in an efficient and cost effective manner withminimal, if any, exposure to biohazards. The sample processing devicesand methods of the present invention are particularly suited to theclinical setting, allowing the clinician to readily proceed fromacquisition of a test sample to analysis of the test results, withminimal human intervention. The sample processing devices of the presentinvention may be implemented as a hand-held system suitable for theprocessing of a single sample or as a larger, bench top unit suitablefor the simultaneous processing of multiple samples. The presentinvention may be valuable in all diagnostic and therapeutic monitoringareas, including in the point-of-care or clinical setting, inhigh-throughput screening, and in biological warfare detection. Inaddition, the present invention provides a sample vessel for holding abiological sample throughout the processing of the sample.

In accordance with one embodiment of the present invention, a device forprocessing a sample includes a processing unit having an opening toreceive a sample vessel and at least one processing station positionedalong the opening. The processing station includes a compression memberadapted to compress the sample vessel within the opening and therebydisplace a content of the sample vessel within the sample vessel. Thecontent displaced by the compression member can be, for example, thesample, a reagent, or a mixture of the content and a reagent

In accordance with another aspect, the processing station may include anenergy transfer element for transferring energy to or from the contentwithin the sample vessel and a control system coupled to the energytransfer element to control the energy transferred to or from thecontent. The energy transfer element can be, for example, an electronicheat element, a microwave source, a light source, an ultrasonic sourceor a cooling element.

In accordance with a further aspect, the energy transfer elementtransfers thermal energy to or from the content within the samplevessel. An energy insulator may be positioned adjacent the processingstation. The energy insulator can be, for example, an energy shieldinglayer, an energy absorption layer, an energy refraction layer, or athermal insulator, depending on the type of energy transfer elementemployed. A temperature sensor may be coupled to the control system tomonitor temperature at the processing station. Alternatively, theprocessing station may include a heat sink to dissipate thermal energyfrom the processing station.

In accordance with another aspect, the processing station may include astationary member opposing the compression member across the opening.The compression member can operate to compress the sample vessel againstthe stationary member within the opening.

In accordance with a further aspect, a driver may be coupled to thecompression member to selectively move the compression member andthereby compress the sample vessel within the opening. The driver canbe, for example, a motor coupled to the compression member by a cam.Alternatively, the driver can be an electromagnetic actuating mechanism.

In accordance with another aspect, the processing device can include asensor for detecting a signal from the content within the sample vessel.An energy source can optionally be provided for applying energy to thecontent within the sample vessel to generate a signal from the content.In one embodiment, the processing device can include an electrophoresissystem comprising a pair of electrodes adapted to have a predeterminedvoltage difference and an electrode actuator for inserting theelectrodes into the sample vessel.

In accordance with a further aspect, the processing device may include areagent injector cartridge actuator adapted to receive a reagentinjector cartridge having at least one needle in fluid communicationwith a reagent reservoir. The reagent injector cartridge actuator can beoperable to move the reagent injector cartridge to inject a quantity ofreagent into the sample vessel.

In accordance with another embodiment of the invention, a sample vesselfor holding a sample includes a sample containing portion for holdingthe sample and a handling portion for handling the sample vessel. Thesample containing portion can have a wall constructed of a flexiblematerial permitting substantial flattening of a selected segment of thesample containing portion. The handling portion can be coupled to thesample containing portion and preferably has a generally rigidconstruction to facilitate handling of the sample vessel.

In accordance with another aspect, the sample containing portion of thesample vessel can be a tubule.

In accordance with a further aspect, the sample vessel can include atleast one pressure gate disposed within the sample containing portion todivide the sample containing portion into a plurality of segments. Atleast one of the segments of the sample vessel can have a filtercontained therein that is structured to separate selected components ofa sample material from other components of the sample material.Additionally, at least one of the segments of the sample vessel cancontain a reagent. The reagent can be, for example, an anticoagulant, acell lyses reagent, a nucleotide, an enzyme, a DNA polymerase, atemplate DNA, an oligonucleotide, a primer, an antigen, an antibody, adye, a marker, a molecular probe, a buffer, or a detection material. Thesample containing portion also can include an electrophoresis segmentcontaining a gel for electrophoresis. The electrophoresis segment caninclude a pair of electrodes adapted to maintain a predetermined voltagedifference therebetween. Additionally, one of the segments can containmultilayer membranes or a micro-array bio-chip for analyzing the sample.

In accordance with another aspect, the sample containing portion caninclude a self-sealing injection channel formed therein. The selfsealing injection channel is preferably normally substantially free ofsample material and capable of fluid communication with the samplematerial in the sample containing portion.

In accordance with another aspect, the sample vessel can include aninstrument for obtaining a sample coupled to the sample vessel.

In accordance with a further aspect, the handling portion of the samplevessel includes an opening for receiving a sample. The sample vesselalso can include a closure for selective closing the opening.Preferably, the closure seats against the handling portion to close theopening. In addition, the instrument for obtaining a sample can becoupled to the closure of the sample vessel.

In accordance with another aspect, the handling portion has a wallthickness greater than a thickness of the wall of the sample containingportion. Preferably, the thickness of the wall of the sample containingportion is less than or equal to 0.3 mm. In one embodiment, the handlingportion can include a cylindrical sleeve sized and shaped to fit over aportion of the sample containing portion. The handling portion ispreferably positioned longitudinally adjacent the sample containingportion.

In accordance with another embodiment, a sample vessel for holding asample includes a sample containing portion having at least one pressuregate disposed within the sample containing portion to divide the samplecontaining portion into a plurality of segments. Preferably, at leastone segment of the sample containing portion has a wall constructed of aflexible material permitting substantial flattening of the segment ofthe sample containing portion.

In accordance with another embodiment, a method of processing a samplewithin a sample vessel includes the steps of introducing the samplevessel into a device for processing the sample and compressing thesample vessel to move the sample within the sample vessel from a firstsegment to a second segment of the sample vessel.

In accordance with another aspect, the method of processing a sample caninclude the step of introducing a reagent to the sample within a segmentof the sample vessel.

In accordance with a further aspect, the method of processing a samplecan include the step of heating the sample in the first segment to afirst temperature. The method can also include the step of heating thesample to a second temperature in the second segment. In one embodiment,the first temperature can be effective to denature the sample and thesecond temperature is one at which nucleic acid annealing and nucleicacid synthesis can occur. The method of processing a sample can furtherinclude the steps of compressing the sample vessel to move the samplewithin the sample vessel from the second segment to the first segment ofthe sample vessel and heating the sample to the first temperature in thefirst segment.

In accordance with another aspect, the method of processing the samplecan include the step of analyzing the sample by detecting a signal fromthe sample within a segment of the sample vessel and analyzing thedetected signal to determine a condition of the sample. The analyzingstep can include applying an excitation energy to the sample within thesegment of the sample vessel. Additionally, the analyzing step caninclude conducting electrophoresis analysis of the sample by applying aselective voltage to the sample within a segment of the sample vessel,detecting light emitted from the sample, and analyzing the detectedlight to determine a condition of the sample.

Alternatively, the analyzing step can include applying an excitationenergy to a bio-array member contained within a segment of the samplevessel, detecting light emitted from the bio-array member, and analyzingthe detected light to determine a condition of the sample. The bio-arraymember can be, for example, a multi-layer membrane or a micro-arraybio-chip.

In accordance with a further aspect, the method of processing a samplecan include the step of agitating the sample within a segment of thesample vessel.

In accordance with another embodiment, a method of treating a samplewithin a sample vessel can include the steps of introducing the samplevessel into a device for processing the sample within the sample vesseland compressing one of the segments to mix the reagent with the samplewithin the sample vessel. Preferably, the sample vessel has a pluralityof segments including a segment for containing a reagent and a segmentfor containing the sample.

In accordance with another aspect, the method of processing the samplecan include the step of introducing the reagent into a reagent segmentof the sample after the step of introducing the sample vessel into thedevice for processing the sample.

In accordance with another embodiment, a thermal cycler includes aprocessing unit having an opening to receive a sample vessel containinga sample. The processing unit can have a first processing station, asecond processing station, and a third processing station positionedalong the opening. The first processing station can include a firstcompression member adapted to compress the sample vessel within theopening and a first energy transfer element for transferring energy tothe sample at the first processing station. The second processingstation can include a second compression member adapted to compress thesample vessel within the opening and a second energy transfer elementfor transferring energy to the sample at the second processing station.The third processing station can include a third compression memberadapted to compress the sample vessel within the opening and a thirdenergy transfer element for transferring energy to the sample at thethird processing station. Compression of the sample vessel by of one ofthe compression members can displace the sample within the sample vesselbetween the processing stations.

The present disclosure is also directed to sample vessels that canpermit the collection and the processing of biological and chemicalsamples, such as, for example, blood, saliva, tissue, or urine, in aclosed system. Sample devices disclosed herein may provide a uniformsample handling system that simplifies the sample collection process andreduces exposure to biohazards. One or more of the sample vesselsdisclosed herein can accommodate multiple fluid samples and a pluralityof assays of different types, while concomitantly reducing the volume ofsample necessary for testing.

In accordance with one exemplary embodiment, a sample vessel maycomprise a tubule having an opening for receiving a sample material andat least one compressible section, a generally rigid container receivingat least a portion of the tubule, and an interface in fluidcommunication with the opening in the tubule. The at least onecompressible section may have a wall constructed at least partially froma material having sufficient flexibility to permit compression ofopposed sections of the wall into contact. The interface may facilitatedelivery of a sample material to the tubule through the opening.

In accordance with another exemplary embodiment, a sample vessel maycomprise a tubule having a plurality of lumens and a wall constructed atleast partially from a material having sufficient flexibility to permitcompression of opposed sections of the wall into contact with oneanother, and a pressure gate connecting at least two lumens of theplurality of lumens. The pressure gate may permit selective fluid flowbetween the at least two lumens.

In accordance with another exemplary embodiment, a sample vessel maycomprise a tubule having a wall that forms a lumen when the tubule is inan open configuration. The wall may have a plurality of sectionsincluding at least a first section of the wall having sufficientflexibility to permit compression of a portion of the tubule and atleast a second section of the wall having sufficient rigidity to supporta flow channel within the tubule during compression of the tubule.

In accordance with another exemplary embodiment, an apparatus fordrawing a sample into a sample vessel may comprise a cylindrical housinghaving an opening for receiving the sample vessel, first means forcompressing a first portion of the sample vessel, and second means forcompressing a second portion of the sample vessel. The first compressionmeans may be positioned at a proximal end of the housing and the secondcompression means may be positioned at a distal end of the housing.

BRIEF DESCRIPTION OF THE DRAWINGS

These and other features and advantages of the sample vessels andprocessing devices and methods disclosed herein will be more fullyunderstood by reference to the following detailed description inconjunction with the attached drawings in which like reference numeralsrefer to like elements through the different views. The drawingsillustrate principles of the sample vessels and methods disclosed hereinand, although not to scale, show relative dimensions.

FIG. 1 is a schematic diagram of a device for processing a sampleaccording to the present invention;

FIG. 2 is a schematic diagram of the device of FIG. 1, illustrating acompression member of a processing station of the device compressing thesample vessel;

FIG. 3 is a schematic diagram of an alternative embodiment of a devicefor processing a sample according to the present invention;

FIG. 4 is a schematic diagram of an alternative embodiment of a devicefor processing a sample according to the present invention;

FIG. 5 is a perspective view of an embodiment of a hand held device forprocessing a sample according to the present invention;

FIG. 6 is a perspective view of an embodiment of a bench top device forprocessing a sample according to the present invention;

FIG. 7 is a perspective view of the device of FIG. 6, illustrating thedevice with the top cover removed;

FIG. 8 is a perspective view of an embodiment of a thermal cyclingprocessing unit according to the present invention;

FIG. 9 is a perspective view of the processing unit of FIG. 8;

FIG. 10 is a partially exploded, perspective view of a processingstation of the processing unit of FIG. 8, illustrating a heat block unitand an insulator block unit of the processing station;

FIG. 11 is a partially exploded, perspective view of the processing unitof FIG. 8, illustrating a plurality of heating block units and insulatorblock units;

FIG. 12 is a partially exploded, perspective view of a processingstation of an alternative embodiment of a processing unit according tothe present invention;

FIGS. 13A-13G are side elevational views, in cross-section, of aprocessing unit of the present invention, illustrating the operation ofthe processing unit;

FIG. 14 is a side elevational view, in cross section, of a gelelectrophoresis analysis unit of the present invention;

FIGS. 15A-15B are side elevational views, in cross-section, ofembodiments of a sample vessel according to the present invention;

FIG. 16 is a side elevation view, in cross section, of a portion of asample vessel according to the present invention, illustrating aninjection channel formed in the sample vessel;

FIG. 17 is a side elevational view of a reagent cartridge according tothe present invention;

FIG. 18 is a side elevational view, in cross-section, of a sample vesselaccording to the present invention;

FIGS. 19A-19C illustrate an alternative embodiment of a processing unitof the present invention.

FIG. 20A is a perspective view of an exemplary embodiment of a samplevessel;

FIG. 20B is a side-elevational view in cross-section of the samplevessel of FIG. 20A;

FIG. 20C is an exploded view of the sample vessel of FIG. 20A,illustrating the tubule and collar removed from the container;

FIG. 21A is a perspective view of an exemplary embodiment of a samplevessel;

FIG. 21B is a side-elevational view in cross-section of the samplevessel of FIG. 21A;

FIG. 21C is an exploded view of the sample vessel of FIG. 21A,illustrating the tubule and collar removed from the container;

FIGS. 22A-22B are side-elevational views in cross-section of anexemplary embodiment of a sample vessel having a pair of lumensseparated by a pressure gate;

FIG. 23 is a side-elevational view in cross-section of an exemplaryembodiment of a sample vessel having three lumens separated by a pair ofpressure gates;

FIG. 24 is a side-elevational view in cross-section of another exemplaryembodiment of a sample vessel having three lumens separated by a pair ofpressure gates, illustrating a self-sealing, reinforced wall section forfacilitating injection by a needle;

FIG. 25A is a perspective view of an exemplary embodiment of a samplevessel having a pair of lumens connected by a micro-fluidic channel;

FIGS. 25B-25C are digital photographs of the sample vessel of FIG. 25Aillustrating fluid flow through the lumens of the sample vessel;

FIG. 26A is a perspective view of an exemplary embodiment of a segmentedsample vessel having a plurality of lumens;

FIGS. 26B and 26C are cross-sectional views of the sample vessel of FIG.26A;

FIG. 27 a perspective view of another exemplary embodiment of asegmented sample vessel having a plurality of lumens, illustrating ahinged cover for the sample vessel;

FIG. 28 a perspective view of an exemplary embodiment of a segmentedsample vessel having a plurality of lumens, illustrating alternativeinterfaces for the sample vessel;

FIG. 29A is a side elevational view in partial cross-section of anexemplary embodiment of a sample vessel, illustrating the compression ofthe sample vessel;

FIG. 29B is a cross-sectional view of the sample vessel of FIG. 29Ataken along a line transverse to the longitudinal axis of the tubule1200;

FIGS. 30A-30C are side elevational views in cross-section of anexemplary embodiment of a sample vessel, illustrating compression of thesample vessel into a plurality of configurations;

FIG. 31 is a side elevational view in cross-section of an exemplaryembodiment of a sample vessel having a composite cross-section and amicro-fluidic flow channel;

FIGS. 32A-32B are side elevational views in cross-section of anotherexemplary embodiment of a sample vessel having a composite cross-sectionand a micro-fluidic flow channel, illustrating the sample vessel in aopen configuration (FIG. 32A) and a compressed configuration (FIG. 32B);

FIG. 33A is a side elevational view in cross-section of an exemplaryembodiment of a sample vessel having a plurality micro-fluidic flowchannels interconnecting a plurality of depressions formed on aninterior wall surface of the sample vessel;

FIG. 33B is a top view of an interior wall surface of the sample vesselof FIG. 33A;

FIGS. 34A and 34B are side elevational views in cross-section of anexemplary embodiment of a sample vessel having a composite cross-sectionincluding opposed planar wall sections, illustrating the sample vesselin an open configuration (FIG. 34A) and a compressed configuration (FIG.34B);

FIG. 35A is a perspective view of an exemplary embodiment of a samplevessel having an adapter for facilitating handling of the sample vesseland/or connecting of the sample vessel to an external device;

FIGS. 35B-35E are side elevational views in cross-section of a pluralityof exemplary embodiments of an adapter connected to the sample vesselillustrated in FIG. 35A;

FIGS. 36A-36E are perspective views of an apparatus for drawing a sampleinto a sample vessel, illustrating the operation of the apparatus; and

FIG. 37 is a perspective view of another exemplary embodiment of asample vessel, illustrating the sample vessel with a portion of the wallremoved to show a microarray on an interior surface of the wall of thesample vessel.

DETAILED DESCRIPTION

To provide an overall understanding, certain exemplary embodiments willnow be described; however, it will be understood by one of ordinaryskill in the art that the sample vessels and methods described hereincan be adapted and modified to provide devices and methods for othersuitable applications and that other additions and modifications can bemade without departing from the scope of the present disclosure.

Unless otherwise specified, the exemplary embodiments described belowcan be understood as providing exemplary features of varying detail ofcertain embodiments, and therefore, unless otherwise specified,features, components, modules, and/or aspects of the exemplaryembodiments can be otherwise combined, separated, interchanged, and/orrearranged without departing from the scope of the present disclosure.Additionally, the shapes and sizes of components are also exemplary andunless otherwise specified, can be altered without affecting thedisclosed devices or methods.

The present disclosure provides devices and methods for processing asample. The term “processing” as used herein generally refers to thepreparation, treatment, analysis, and/or the performance of othertesting protocols or assays on a content of the sample vessel in one ormore steps. Exemplary processing steps include, for example: displacinga content, e.g., the sample or a reagent, of the sample vessel withinthe sample vessel to, for example, adjust the volume of the content,separate content components, mix contents within the sample vessel;effecting a chemical or biological reaction within a segment of thesample vessel by, for example, introducing a reagent to the sample,agitating the sample, transferring thermal energy to or from the sample,incubating the sample at a specified temperature, amplifying componentsof the sample, separating and/or isolating components of the sample; oranalyzing the sample to determine a characteristic of the sample, suchas, for example, the quantity, volume, mass, concentration, sequence, ornucleic acid size or other analyte size, of the sample. One skilled inthe art will appreciate that the forgoing exemplary processing steps aredescribed herein for illustrative purposes only. Other processing stepsmay be employed without departing from the scope of the presentinvention.

A device for processing a sample according to the present invention canintegrate one or more processing units into a single system depending onthe process being employed. The processing units can include one or moreprocessing stations at which one or more processing steps can beperformed on the sample within the sample vessel. Sample materials thatcan be processed according to the present invention are generallybiological samples or samples containing biological substance andinclude, for example, blood, urine, saliva, cell suspensions, biofluids,a piece of tissue, soil or other samples. A sample processing device ofthe present invention is particularly suited for nucleic acidamplification, such as polymerase chain reaction (PCR) or ligase chainreaction (LCR) amplification, and can include, for example, a samplepretreatment unit for extracting nucleic acid from sample, a thermalcycling reaction unit for amplification of the nucleic acid or signal,and (optionally) an analysis or detection unit for analyzing theamplified product. The sample processing device of the present inventioncan also be used for isothermal reaction of nucleic acid or signalamplifications, such as strand displacement amplification (SDA), rollingcircle amplification (RCA), and transcription-mediated amplification(TMA). Other exemplary processes to be performed on samples can includeclinical diagnosis, therapeutic monitoring, and screening of chemicalcompounds for discovery of new drugs. The following descriptionprimarily focuses on PCR amplification for illustration. However, oneskilled in the art will appreciate that the devices and methods of thepresent invention are not limited to PCR amplification, as the devicesand methods described below can be employed in other sample processing.

An exemplary embodiment of a device for processing a sample isillustrated in FIG. 1. The processing device 10 illustrated in FIG. 1includes a processing unit 12 having an opening 14 to receive a samplevessel 16. The opening 14 can be a tubular shaped opening, an open-facedslot or other structure for receiving the sample vessel 16 in aremovable and replaceable manner. The processing unit 12 includes afirst processing station 18 and a second processing station 20, eachpositioned along the length of the opening 14. The first processingstation 18 includes a compression member 22 adapted to compress thesample vessel 16 within the opening 14 and thereby displace a content ofthe sample vessel within the sample vessel 16. The content of the samplevessel can be, for example, the sample, a reagent contained within thesample vessel, or a mixture of the sample and the reagent. A driver 24is coupled to the compression member 22 to selectively move thecompression member 22 and thereby compress the sample vessel 16 withinthe opening 14. The driver 24 can be, for example, an electromagneticactuating mechanism, a motor, a solenoid, or any other device forimparting motion, preferably reciprocal motion, to the compressionmember 22, as described in further detail below.

Preferably, the compression member 22 is constructed from a rigidmaterial such as a rigid plastic or a metal. The compression member canbe constructed in any shape sufficient to impart a compressive force onthe sample vessel. For example, the compression member 22 can be a blockhaving a rectilinear, planar surface for engaging the sample vessel 16,as illustrated in FIG. 1. Alternatively, the compression member can havea curved, angular, or non-planar surface for engaging the sample vessel16.

Moreover, the compression member 22 alternatively can be an inflatablemembrane that can be inflated by a fluid, e.g., air, nitrogen, saline,or water, to impart a compressive force on the sample vessel. In thisembodiment, the amount of compression of the sample vessel may becontrolled by the adjusting the inflation pressure of the membrane.

The first processing station 18 can optionally include a stationarymember 26 positioned opposite the compression member 22 across theopening 14. The compression member 22, thus, can compress a portion ofthe sample vessel 16 within the opening 14 against the stationary member26, as illustrated in FIG. 2. One skilled in the art will appreciatethat the stationary member 26 may be replaced with a second compressionmember, such that the processing station includes two compressionmembers that move together to compress the sample vessel therebetween.In addition, a stationary member or second compression member may beomitted by securing the sample vessel 16 within the opening on eitherside of the compression member.

In the illustrated embodiment, the sample vessel 16 is a closed tubuleflow-chamber for holding the sample. Preferably, one or more segments ofthe sample vessel 16 are constructed of a flexible, compressiblematerial, such as, for example, polyethylene or polyurethane, to allowselective compression, and preferably flattening, of the sample vesselto move the sample, or other contents of the sample vessel, within thesample vessel, preferably while the sample vessel 16 remains in thedevice 10. In one preferred embodiment, the sample vessel 16 includes aplurality of segments separated by an integral, internal structure, suchas a micro-fluidic pressure gate, as described in more detail below.Alternatively, the sample vessel 16 can be constructed without internal,integral structures to form segments and the device 10 can be utilizedto segment the sample vessel by compressing selective portions of thesample vessel. One skilled in the art will appreciate that other typesof sample vessels suitable for containing a sample may be used with thedevice 10 without departing from the scope of the present invention.

The second processing station 20 can include a sensor 28 for detecting asignal from the content, e.g., the sample or a reagent, of the samplevessel 16. For example, the sensor 28 can be an optical sensor formeasuring light, for example fluorescent light, emitted from the sampleor from fluorescent probes within the sample. In addition, multiplesensors or a spectrum sensor can be used when detection of multiplewavelength light is required. The detected signal can be sent to a CPU30 to analyze the detected signal and determine a characteristic of thesample.

In operation, a sample can be introduced to a first segment A of thesample vessel 16 by injecting the sample through the walls of the samplevessel 16 or by introducing the sample through an opening formed in thesample vessel 16, as described in more detail below. In the presentexemplary embodiment illustrated in FIGS. 1 and 2, the sample vessel 16includes a pressure gate 32 that divides the sample vessel 16 into afirst segment A and a second segment B. The sample vessel 14 can beinserted into the opening 14 of the device 10 such that the firstsegment A of the sample vessel 16 is aligned with the first processingstation 18 and the second segment B is aligned with the secondprocessing station 20, as illustrated in FIG. 1.

The driver 24 can operate to move the compression member 22 into contactwith the sample vessel 16 such that the first segment A of the samplevessel 16 is compressed within the opening 14 between the compressionmember 22 and the stationary member 26. As the first segment A of thesample vessel 16 is compressed, a quantity of sample is displaced fromthe first segment A to the second segment B through the pressure gate32. The volume of sample displaced is proportional to the amount ofcompression of the first segment A by the compression member 22. Thus,the compression member 22 of the first processing station 18 can be usedto displace a specific quantity of sample into the second segment B ofthe sample vessel 16 for analysis at the second processing station 20.Substantially all of the sample can be displaced from the first segmentA of the sample vessel 16 by completely flattening the first segment Aof the sample vessel 16, as illustrated in FIG. 2. The sample can beanalyzed in the second segment B of the sample vessel 16 at the secondprocessing station 20.

An alternative embodiment of a device for processing a sample isillustrated in FIG. 3. The device 38 includes a processing unit 40having three processing stations positioned along the opening 14,namely, a first process station 42, a second processing station 44adjacent the first processing station 42, and a third processing station46 adjacent the second processing station 44.

The first processing station 42 includes a compression member 22 coupledto a driver 24 and adapted to compress a segment of the sample vessel 16against a stationary member 26 within the opening 16. The firstprocessing station 42 can operate to displace a selective quantity ofthe sample from a first segment A of the sample vessel into othersegments of the sample vessel.

The second processing station 44 includes a compression member 22coupled to a driver 24 and adapted to compress a second segment B of thesample vessel 16 against a stationary member 26 within the opening 16.The second processing station 44 includes an energy transfer element 48for transferring energy to or from the contents of the sample vessel 16.The energy transfer element 48 can be, for example, an electronic heatelement, a microwave source, a light source, an ultrasonic source, acooling element, or any other device for transferring energy. In oneembodiment, the energy transfer element 48 transfers thermal energy toor from the sample within the sample vessel. The energy transfer element48 can be embedded in or otherwise coupled to the compression member 22,as illustrated in FIG. 3. Alternatively, the energy transfer element 48can be coupled to the stationary member 26 or can be positioned withinthe processing station independent of the compression member or thestationary member. The energy transfer element 48 can be coupled to acontrol system that controls the energy transferred to or from thesample vessel 16 by the energy transfer element 48. The control systemcan be a component system of the CPU 30 or can be an independent system.The control system can also include a temperature sensor 50 to monitorthe temperature of the energy transfer element.

The second processing station 44 also can include a sensor 52 fordetecting a signal from the content of the sample vessel, particularlyduring processing in the second processing station. For example, thesensor 52 can be an optical sensor for measuring light, for examplefluorescent light, emitted from the sample or from fluorescent probeswithin the sample. The sensor 52 can be coupled to the CPU 30 foranalysis of the detected signal to determine a characteristic of thesample.

The third processing station 46 can include a sensor 28 for detecting asignal from the content, e.g., the sample or a reagent, of the samplevessel 16. For example, the sensor 28 can be an optical sensor formeasuring light, for example fluorescent light, emitted from the sampleor from fluorescent probes within the sample. In addition, multiplesensors or a spectrum sensor can be used when detection of multiplewavelength light is required. The detected signal can be sent to a CPU30 to analyze the detected signal and determine a characteristic of thesample.

In operation, a sample can be introduced into a first segment A of thesample vessel 16 and the sample vessel 16 can be introduced into theopening 14 of the device 10. In the embodiment illustrated in FIG. 3,the sample vessel 16 includes two pressure gates 32 that divide thesample vessel 16 into three segments, namely, the first segment A, asecond segment B, and a third segment C. The first processing station 42can operate to displace a selective amount of the sample into the secondsegment B of the sample vessel 16 for processing at the secondprocessing station 44.

At the second processing station 44, energy can be transferred to orfrom the sample within the second segment B. In this manner, abiological or chemical reaction involving the sample may be carried outin the second segment B. The sensor 52 can be used to monitor thereaction during the reaction process.

Upon completion of the reaction, the sample can be moved into the thirdsegment C of the sample vessel 16 by compressing the sample vessel 16within the opening at the second processing station 44. Preferably, thecompression member 22 of the first processing station 42 substantiallyflattens the first segment A of the sample vessel 16 to inhibit thesample from entering the first segment A. The sample can be analyzed inthe third segment C of the sample vessel 16 at the third processingstation 46.

A further embodiment of a device for processing a sample is illustratedin FIG. 4. The device 56 includes a processing unit 58 having aprocessing station 60 positioned along the opening 14. The processingstation 60 includes a compression member 22 coupled to a driver 24 andadapted to compress a segment of the sample vessel 16 against astationary member 26 within the opening 16. In the embodimentillustrated in FIG. 4, the sample vessel 16 includes a pressure gate 32that divides the sample vessel 16 into two segments, namely, a firstsegment A and a second segment B. The processing station 60 can operateto displace a selective quantity of the content from the second segmentB of the sample vessel into the first segment A of the sample vessel.For example, a reagent can be introduced into the second segment B ofthe sample vessel 16. A quantity of reagent can be displaced from thesecond segment B into the first segment A of the sample vessel 16 to mixwith the sample in the first segment A. Alternatively, the reagent canbe introduced into the first segment A of the sample vessel 16 and aquantity of the sample can be displaced from the second segment B intothe first segment A by the processing station 60. Thus, the firstsegment A of the sample vessel 16 can act as a reaction mixture chamberfor the sample and the reagent. The reagent can be pre-packaged in thesample vessel 16 or can be introduced to the sample vessel 16 after thesample is introduced to the sample vessel 16. For example, the reagentcan be introduced using a reagent injector cartridge, described below,that is included with the device.

Referring to FIG. 5, another embodiment of device for processing asample is illustrated. The illustrated device 100 is a hand held systemfor processing a nucleic acid sample, preferably in an “insert and test”format in which a sample vessel containing a nucleic acid sample isinserted into the device 100 and processing results are produced by thedevice with minimal human intervention. The device 100 can include ahousing 112 having an opening 114 for receiving a sample vessel 116containing a sample for processing by the device 100. The opening 114can be a tubular shaped opening, as illustrated in FIG. 5, or can be anopen-faced slot or other structure for receiving the sample vessel in aremovable and replaceable manner. A control panel 118 is located on thetop of the housing 112 for inputting information to the device 100 and amonitor 120 is provided for displaying operating information, such asthe results of processing. An external communication port 121 can belocated on the housing 112 for receiving information or outputtinginformation, such as the results of processing and remote diagnosing ofthe system, to a remote system, such as a computer network. A battery123 (FIG. 7) can be located within the housing to provide electricalpower to the components of the device 100.

A multi-sample device 200 for processing multiple samples is illustratedin FIG. 6. The device 200 is a bench top thermal cycling system forprocessing up to 96 nucleic acid samples simultaneously. The sampleprocessing device 200 operates on the same principals as the sampleprocessing device 100 illustrated in FIG. 5, except that themulti-sample device 200 provides increased capacity and throughput. Themulti-sample processing device 200 can include a housing 202 having aplurality of wells or openings 204, with each well being capable ofreceiving a sample vessel 206 containing a sample for processing by thedevice. The exemplary multi-sample device 200 illustrated in FIG. 6 hasninety-six wells for treating up to 96 samples simultaneously. Oneskilled in the art will appreciate that a multi-sample processing deviceaccording to the present invention may be designed with any number ofwells, depending on the sample being tested and the processes beingemployed, without departing from the scope of the present invention. Acontrol panel 208 is located on the top of the housing 202 for inputtinginformation to the multi-sample processing device 200 and a monitor 210is provided for displaying operating information, such as the results oftesting.

FIG. 7 illustrates the general components of the sample processingdevice 100 illustrated in FIG. 5. The illustrated device 100 includesthree primary processing units for processing a sample within the samplevessel, namely, a pretreatment unit 122 for pretreating the sample, areaction unit 124 for amplifying certain components of the sample, andan analysis unit 126 for analyzing the sample. The sample vessel can beloaded into the device 100 through the opening 114. The processing unitsof the device are preferably arranged along the axis of elongation ofthe opening 114. This arrangement allows the sample to be moved withinthe sample vessel between the processing units of the device 100 in amanner described in detail below. Preferably, the processing units arearranged linearly as illustrated in FIG. 7, however, other arrangementare possible so long as the sample vessel can be positioned adjacent oneor more of the processing units of the device 100.

Continuing to refer to FIG. 7, a pair of sample vessel loading wheels128 is located at the entrance 130 of the sample vessel opening 114. Theentrance 130 is preferably tapered to facilitate loading of the samplevessel into the opening 114 of the device 100. The loading wheels 128further facilitate loading of the sample vessel by guiding the samplevessel into the opening 114. A sample collection unit 132 can bepositioned proximate the entrance 130 of the opening 114 to allow aselective volume of the sample to dispense into the next processing unitor units within the sample vessel. The sample collection unit 132 caninclude a compression member 22 opposed to a stationary member 26 acrossthe width of the opening 114. A linear motor 138 is coupled to thecompression member 22. The linear motor 138 can operate to move thecompression member 22 toward or away from the stationary member 26 toselectively open and close the opening 114 therebetween. When the samplevessel is positioned within the opening 114, the linear motor 138 canoperate to compress the sample vessel between the compression member 22and the stationary member 26. As a result, a selective volume of thesample can be moved to the next processing unit within the samplevessel. Preferably, the sample vessel remains compressed between thecompression member 22 and the stationary member 26 of the samplecollection unit 132 during processing of the sample by the otherprocessing units to prevent the sample from exiting the processing unitarea during processing.

The pretreatment unit 122 is positioned adjacent the initial samplecollection unit 132. Depending on the process being implemented, thesample may require pretreatment or preparation before proceeding withadditional processing steps. Pretreatment can include, for example,adding a reagent or other material to the sample and incubating themixture for certain time period. The pretreatment unit 122 of the device100 allows for any of such pretreatment steps to be implemented. For PCRtesting, the sample pretreatment unit 122 can provide for nucleic acidextraction from a biological sample, such as blood. Any known methodsfor extracting nucleic acid can be utilized in the pretreatment unit,including using a cell lysis reagent, boiling the nucleic acid sample,GITC, or formamide for solubilization. Alternatively, filters can beused within the sample vessel to separate nucleic acid from unwantedcellular debris.

The pretreatment unit 122 can include a compression member 22 and astationary member 26 opposed to the compression member 26 across theopening 114. The compression member 22 and/or the stationary member 26can optionally include an energy transfer element for transferringenergy, e.g. thermal energy, to the sample within the sample vessel. Theenergy transfer element can be, for example, an electronic heat element(such as Kapton heater, a Nomex heater, a Mica heater, or a siliconerubber heater), a microwave generator, a light source, an electroniccooling element (such as Peltier element), an ultrasonic energy transferelement, or any another device suitable for transferring thermal energy.A driver 24, for example an electromagnetic actuator such as linearstepper actuator, a relay actuator, or a solenoid, is coupled to thecompression member 22 and operates as a driver. During operation of thepretreatment unit 122, the driver 24, moves the compression member 22 toopen the opening 114 between the compression member 22 and thestationary member 26 of the pretreatment unit 122 to allow receipt of asample vessel. After a sample vessel is loaded, the driver 24 drives thecompression member 22 toward the stationary member 26, resulting in goodsurface contact between the sample vessel and the compression member andthe stationary member and thus improved pretreatment. Once thepretreatment is completed, the driver 24 moves the compression member 22of the pretreatment unit 122 to further compress the pretreatmentsegment of the sample vessel to move a selective amount of pretreatedsample within the sample vessel to the next processing unit.

The reaction unit 124 can include a plurality of processing stations150A-150C and is preferably positioned adjacent the pretreatment unit122. The reaction unit 124 can effect thermal cycling of the sample byselectively moving the sample, with the sample vessel, between theprocessing stations 150A-150C. The phrase “thermal cycling” as usedherein refers to a process of heating and/or cooling a sample in two ormore steps, with each step preferably occurring at a differenttemperature range from the previous step. Each of the processingstations 150A-150C can be maintained at a pre-selected temperature rangecontrolled by a temperature control system 152 and a CPU 174. Althoughthe exemplary embodiment includes three thermal cycling processingstations 150A-150C, the reaction unit 124 can include any number ofprocessing stations 150, depending on the thermal cycling processemployed. Alternatively, the reaction unit 124 can incubate a sample ata selective temperature for an isothermal reaction such as for TMA orSDA process.

In PCR based testing, thermal cycling can be used to denature, anneal,elongate and thereby amplify the nucleic acid sample. The PCR thermalcycling steps each occur at specified temperature ranges. Denaturingoccurs at approximately 92° C.-96° C.; elongation occurs atapproximately 70° C.-76° C.; and annealing occurs at approximately 48°C.-68° C. Each of the PCR thermal cycling steps, i.e. denaturing,annealing, and elongation, can be carried out independently at aseparate processing station of the reaction unit 124 by maintaining theprocessing stations at the temperature ranges effective for carrying outeach of the PCR thermal cycling steps. For example, the denaturing stepcan be carried out at processing station 150A, the elongation step atprocessing station 150B, and the annealing step at processing station150C. Alternatively, one or more of the PCR thermal cycling steps can becombined and carried out at the same processing station, therebyreducing the number of processing stations required. For example,denaturing can be carried out at processing station 150A and elongationand annealing can be carried out at processing station 150B, thus,eliminating the need for a third processing station.

Moreover, a processing station can be provided within the reaction unit122 for cooling of the sample by using a thermal energy element, aPeltier thermal electric element for example, to transfer thermal energyfrom the processing station. In PCR processing, for example, aprocessing station can be provided to preserve the nucleic acid samplebetween process steps by cooling the sample to a refrigerationtemperature, e.g., 4° C. Additionally, a processing station canoptionally be provided to cool the sample between thermal cycling stepsto decrease the temperature down ramping time between successive thermalcycling steps. For example, as denaturing generally occurs at 92° C.-96°C. and annealing generally occurs at a significantly lower temperature,e.g., 48° C.-68° C., the sample can be cooled after denaturing in acooling processing station, preferably at a temperature lower than theannealing temperature, to bring the sample temperature more quickly intothe annealing temperature range. A thermal cycling processing stationcan optionally include a heat sink 166 coupled to either the compressionmember 22 or the stationary member 26 to conduct heat away from thestation and radiate the heat to the environment.

Each of the illustrated processing stations of the reaction unit 124includes a compression member 22 and a stationary member 26. Thecompression member 22 of each thermal cycling processing unit can becoupled to a driver 24 for selectively moving the compression member 22toward and away from the stationary member 26. As discussed above, thedrivers 24 can be any device capable of imparting motion, preferablyreciprocal motion, to the compression members. A driver control system160 is coupled to the drivers 24 to control the operation of the drivers24. In one preferred embodiment illustrated in FIG. 7, the drivers 24are electromagnetic actuators coupled to the driver control system 160,which can be, for example, a control system for controlling thereciprocal motion of the actuators. Alternative drivers, compressionmembers and stationary members are described below in connection withFIGS. 8-12. The driver control system 160 is coupled to the CPU 174 suchthat the sample incubation time period, the pressure and the samplemoving speed within the sample vessel can be controlled and coordinatedby the CPU 174 to achieve the best reaction results.

Each of the thermal cycling processing station 150A-150C can optionallyinclude an energy transfer element for transferring energy, such asthermal energy, to the sample within the sample vessel. The energytransfer elements can be, for example, an electronic heat element, amicrowave generator, a light source, an electronic cooling element, orany another device suitable for applying thermal energy. Each of theenergy transfer elements is coupled to the temperature control system152 to maintain the associated processing station within a selectedtemperature range. One or more temperature sensors, coupled to thetemperature control system 152, can be positioned proximate theprocessing stations 150A-150C to monitor the temperature of thestations.

Between two adjacent processing units or two adjacent processingstations, at least one layer of energy insulator 146 can optionally beprovided to insulate the processing unit or the processing station fromadjacent units or stations. An energy insulator layer can also be formedon the boundary of a processing station to prevent energy transfer to orfrom the environment. The energy insulator 146 can be, for example, anenergy shielding layer, an energy absorption layer, an energy refractionlayer, or a thermal insulator, depending on the type of energy transferelement employed. A thermal insulator can be constructed from a lowthermal conductivity material such as certain ceramics or plastics. Inone embodiment, the thermal insulator can be attached to the compressionmembers and the stationary members. Alternatively, the thermalinsulators can be separate from the compression members and stationarymembers and can be controlled independently by a driver to open andclose the opening 114. In this embodiment, all the compression membersand insulators can open initially to allow loading of the sample vessel,and then, the thermal insulators can compress the sample vessel withinthe opening to close the vessel and form separate segments within thesample vessel. Additionally, a spring element or other biasing mechanismcan be optionally utilized to bias each thermal insulator. Through thespring element, a driver associated with one of the thermal insulatorscan apply sufficient pressure on the thermal insulator to minimize thequantity of sample remaining in the junction between adjacent processingstations during an incubation period, while still allowing sample flowthrough the thermal insulator when a higher pressure is applied to thesample in an adjacent processing station. This design simplifies theoperation of multiple thermal insulators.

In an alternative embodiment, the processing stations can be spacedapart to inhibit conductive heat transfer between adjacent processingstations and, thereby, eliminate the need for insulators between thestations.

Operation of the thermal cycling reaction unit 124 will be generallydescribed with reference to FIGS. 13A-13G. The thermal cycling processbegins by opening each of the processing stations, e.g. first processingstation 150A, second processing station 150B, and third processingstation 150C, to receive the sample vessel within the opening 114, asillustrated in FIG. 13A. After the sample vessel is loaded withpretreated sample material, or the pretreated sample is dispensed frompretreatment unit 122 into the reaction unit 124, the second processingstation 150B and the third processing station 150C are closed by movingthe compression member 22B and the compression member 22C of eachstation toward the respective stationary member 26B and 26C, asillustrated in FIG. 13B. As the second processing station 150B and thethird processing station 150C are closed, the sample vessel iscompressed between the compression member and the stationary member,displacing the sample within the sample vessel into a segment of thesample vessel adjacent the first processing station 150A.

Next, the compression member 22A and the insulator 146A can compress thesample vessel to adjust the sample volume contained within the segmentof the sample vessel adjacent the first processing station 150A, as wellas the surface area to volume ratio of the segment. The insulator 146Acan then be closed to seal the sample in the first processing station150A, as illustrated in FIG. 13C. Alternatively, if the device 100 isprovided with a sample pretreatment unit, the sample pretreatment unitcan function to close the sample vessel within the first processingstation 150A. Other alternatives include pre-sealing the sample vesselafter loading a sample, or providing the sample vessel with pressuregates, discussed below, formed between adjacent reaction zones. Once thesample is sealed within the first processing station 150A, the samplecan be heated or cooled by the first processing station 150A. In PCRthermal cycling, for example, the sample can be heated to a temperatureeffective to denature the nucleic acid sample. Preferably, the samplevessel is pressed into contact with the compression member 22A and thestationary member 26A by the compression member 22A to flatten thesample vessel and to ensure good thermal contact between the samplevessel and the compression member 22A and the stationary member 26A. Thecompression member 22A can also optionally periodically squeeze thesample vessel to agitate the sample and to generate sample flow in thesegment of the sample vessel during the reaction period to speed up thereaction.

After a predetermined period, the second processing station 150B can beopened to allow the sample to move into the second processing station150B, as illustrated in FIG. 13D. Next, the first processing station150A closes, compressing the sample vessel and moving the entire sample,within the vessel 16, into a segment of the sample vessel adjacent thesecond processing station 150B, as illustrated in FIG. 13E. The thirdprocessing station 150C can then open to allow the sample to move intothe segment of the sample vessel adjacent the third processing station150C, as illustrated in FIG. 13F. The second processing station 150Bcloses, compressing the sample vessel and moving the sample completelyinto the segment of the sample vessel adjacent the third processingstation 150C, as illustrated in FIG. 13G. The sample can then be heatedor cooled by the third processing station 150C for a set time period. InPCR thermal cycling for example, the sample can be heated to atemperature effective to anneal the nucleic acid sample in the thirdprocessing station 150C. The heat sink 166 can facilitate thetemperature transition from the denaturing temperature of the firstprocessing station 150A to the annealing temperature of the thirdprocessing station 150C by dissipating excess heat to the environment.Thus, the sample can be moved from the denaturing step at the firstprocessing station to the annealing step at the third processingstation.

After a predetermined time period, the second processing station 150Bopens to allow the sample to move into the second processing station, asillustrated in FIG. 13F. The third processing station 150C then closes,compressing the sample vessel 16 and moving the sample entirely into thesegment of the sample vessel adjacent the second processing station150B, as illustrated in FIG. 13E. The sample can then be heated orcooled by the second processing station 150B for a set time period. InPCR thermal cycling for example, the sample can be heated to atemperature effective to elongate the nucleic acid sample. Uponconclusion of the elongation step, the sample can be returned to thesegment of the sample vessel adjacent the first processing station 150Ato repeat the cycle, i.e., denaturing and annealing and elongating or,the sample can be moved to a segment of the sample vessel adjacent thesample detection unit 126 if PCR thermal cycling is completed.

The illustrated thermal cycling reaction unit 124 provides denaturing inthe first processing station 150A, annealing in the third processingstation 150C, and elongation in the second processing station 150B, asthis arrangement is deemed thermodynamically efficient. One skilled inthe art will appreciate, however, that the PCR thermal cycling steps canoccur in any of the processing stations without departing from the scopeof the present invention.

Sample thermal cycling using the reaction unit 124 of the presentinvention results in faster thermal cycling times and lower energyconsumption compared to conventional thermal cycling devices. Samplevessel shape alteration, i.e. flattening, by the reaction unit 124results in significant increases in the surface/volume ratio and samplevessel contact with the members of the reaction unit. This allows theprocessing stations of the reaction unit 124 to heat the sample moredirectly, increasing the sample temperature ramping rate and avoidingunnecessary temperature ramping of the members and thus decreasing theamount of energy consumed. Equally important is that sample vessel shapealteration provides for the uniform transfer of thermal energy to thesample, dramatically reducing reaction mixture temperature gradients.The reaction unit 124 further allows the use of fluid flow to mix thesample as the sample is moved from one processing station to another.

Moreover, the reaction unit 124 allows the use of a disposable,single-use sample vessel that minimizes contamination of the sample,contamination of the reaction unit and exposure of the operator tobiohazards. Additionally, the reaction unit 124 does not require a fluidhandling system, as the sample can be moved within the sample vesselduring processing.

Referring again to FIG. 7, the reaction unit 124 can optionally includea reaction sensor 168 for monitoring the reaction in real-time withinthe reaction unit 124 by analyzing the sample, including any reactionproducts from the reaction with the sample. The reaction sensor 168 caninclude an integral light source 169 for applying excitation energy tothe sample within the sample vessel. Alternatively, a light source, orother source of excitation energy, can be provided separate from thereaction sensor 168. The reaction sensor 168 can be an optical sensorfor measuring light, for example fluorescent light, emitted from thesample or from fluorescent probes within the sample. In the case of PCR,any known real-time PCR detection system can be employed, including, forexample, using fluorescent dyes, such as ethidium bromide, intercalatinginto the DNA molecule, using a dual labeled probe tagged with a reporteddye and a quenching dye, or using hybridization probes, which willresult in Fluorescence Resonance Energy Transfer (FRET) only when thetwo probes are hybridized and in close proximity. In each of theseapproaches, the fluorescence signal is substantially proportional to theamount of specific DNA product amplified. The reaction detection sensor168 is placed to monitor the fluorescence from the sample, preferably inthe annealing processing station, or other processing stations of thereaction unit, dependent on the assay selected. Multiple sensors or aspectrum sensor can be used when detection of multiple wavelength lightis required. The detected signal is then sent to the CPU 174 for furtheranalyzing the amount of product.

Continuing to refer to FIG. 7, the sample detection or analysis unit 126of the device 100 is provided to analyze the sample after processing bythe reaction unit 124. The analysis unit 126 is preferably positionedproximate the reaction unit 124. In one embodiment of the invention, asource of excitation energy, for example a light source, can applyexcitation energy to the sample and a signal detector, for example anoptical sensor, can detect light emitted from the sample in response toillumination by the excitation light. Specific illustrative practices,include detecting the transmission of light through the sample,detecting reflected light, detecting scattering light, and detectingemitted light. The detected light, in the form of the signal output fromthe sensor, can be analyzed by a CPU 174 provided in the device throughknown signal processing algorithms. Suitable sample analysis systems,employing a light source and an optical sensor or sensors, detectssignals including light intensity at a given wavelength, phase orspectrum of the light, as well as location of the signal. For example,the flow induced testing system described in U.S. Pat. No. 6,318,191 andthe multi-layer testing system described in U.S. Pat. App. Pub. No. US2004/0105782 A1, both of which are incorporated herein by reference,describe suitable sample analysis systems.

In the case of a PCR based assay, gel electrophoresis or capillaryelectrophoresis can be employed to analyze the nucleic acid sample, asillustrated in FIGS. 7 and 14. Suitable nucleic acid sizing gels includeagarose and polyacrylamide. The gel 184 can be introduced to the samplevessel 16 during processing or, preferably, is pre-loaded into ananalysis segment 210 of the sample vessel, as discussed in more detailbelow. The exemplary analysis unit 126 includes a light source 170 forilluminating the nucleic acid sample and the gel and an optical sensor172 in the form of linear charge coupled device (CCD). Electrodeactivators 176 operate to insert a positive electrode 180 and a negativeelectrode 182 into the sample vessel 16. The positive electrode 180 andthe negative electrode 182 are electrically connected to a voltagesource, which creates a voltage difference between the electrodes. Asnucleic acid products are negatively charged, the nucleic acid productswithin the sample will move through the gel 184 toward the positiveelectrode 180. The gel separates the sample components by size, allowingsmaller components, such as nucleic acid products, to travel faster, andthus, further, than larger components. A suitable dye or fluorescent tagcan be introduced to gel to identify the nucleic acid products. Lightfrom the light source 170 can illuminate the dyed or tagged nucleic acidproducts in the gel and the optical sensor 172 can then identify theilluminated nucleic acid products. The output signal of the opticalsensor 172 can be analyzed by CPU 174 according to known signalprocessing method to determine the presence, absence, quantity or othercondition of the nucleic acid sample.

Alternatively, the nucleic acid sample can be analyzed in accordancewith conventional nucleic acid analysis methods, such as, for example,chemiluminescence, fluorescently labeled primers, antibody capture, DNAchip, and/or magnetic bead detection methods.

One skilled in the art will appreciate that the processing units and theprocessing stations of the above-described exemplary embodiments of thesample processing device of the present invention can be arranged in anyorder depending on the sample being processed and the process beingutilized. The sample processing device of the present invention mayinclude any combination of the processing units and processing stationsdescribed herein, as well as additional processing units and processingstations that will be apparent to those skilled in the art upon readingthis disclosure. Moreover, the sample processing device may include onlya single processing unit, such as, for example, a reaction unit forthermal cycling a sample, or may include a only a single processingstation, such as, for example, a processing station for displacing aspecified volume of reagent or sample.

FIGS. 8-12 illustrate alternative embodiments of a reaction unit 250 forthermal cycling a sample according to the present invention. Thereaction unit 250 can include one or more openings 252 for receiving oneor more sample vessels 16. The embodiments illustrated in FIGS. 8-12have three openings 252, permitting the simultaneous thermal cycling ofup to three samples. The reaction unit 250 comprises three processingstations: a first processing station 254, a second processing station256, and a third processing station 258. Thermal insulators 260A-260Dare positioned between the processing stations and at the top of thefirst processing station 254 and the bottom of the third processingstation 258.

Referring specifically to FIGS. 8 and 10, the first processing station254, as well as the second and third processing stations 256 and 258,includes an embedded heat element 262 for transferring thermal energy tothe sample vessel when the sample vessel is positioned within an opening252. The heat element 262 can be a Kapton heater, a Nomex heater, a Micaheater, a silicone rubber heater or any other thermal energy transferelement suitable for delivering thermal energy. The heat element 262 canbe seated in a recess 264 formed in the processing station 254 andsecured to the processing station by an adhesive or other attachmentmeans. The heat element 262 of each of the processing stations ispreferably coupled to a temperature controller 266 for controlling thetemperature of the heat element. One or more temperature sensors 268 canbe positioned in the processing station 254 to measure the temperatureof the processing station 254. The temperature sensor 268 can be coupledto the thermal controller 266 such that the temperature controller 266can monitor and adjust the temperature of the processing station in afeedback control manner.

Referring to FIGS. 10 and 11, each processing station comprises astationary member 270 and a compression member 272 adapted to compressthe sample vessel selectively within one or more of the openings 252 andthereby move the sample within the sample vessel. The compression member272 is preferably complimentary in shape to the stationary member 270and includes a plurality of finger-like closure elements or shutters 274sized and shaped to slide within the openings 252. Guide rails 276 arepositioned on either side of the compression member. The guide rails 276are preferably sized and shaped to fit within grooves 278 formed in theside walls of the stationary member 270. The combination of the guiderails 276 and the grooves 280 allow the compression member 272 toreciprocate relative to the stationary member 270 to selectively openand close the openings 252.

Each thermal insulator 260 can be configured in a manner analogous tothe processing stations. For example, the thermal insulator 260Bcomprises an insulator stationary member 280 and an insulatorcompression member 282 adapted to compress a sample vessel within one ormore of the openings 252. The insulator compression member 282 includesa plurality of finger-like closure elements or shutters 284 sized andshape to slide within the openings 252 to selectively open and close theopenings 252.

Each compression member 272 and insulator compression member 282 iscoupled to a driver, such as an electromagnetic driver mechanism, asdescribed above, or any other mechanism for imparting motion, preferablyreciprocating motion, to the compression members. Each compressionmember can be coupled to an arm 286 for providing a connection betweenthe compression member and the driver, as best illustrated in FIGS.9-11. In one embodiment, illustrated in FIG. 10, the arms 286 are hollowtubes that receive coiled springs 288 and dowels 290. The springs 288operate to bias the compression members 272, 282 in a direction awayfrom the stationary member 270 and the insulator stationary member 280,respectively. An elastic element, such as the coiled spring used here,provides a simple mechanism for assisting the driver to regulate thecompressing pressure applied to the sample vessel. The driver can be amotor 292 for driving a rotating shaft, as best illustrated in FIG. 8.The rotary motion of the shaft can be translated to reciprocating motionthrough cams 294 provided for each of the compression members 272 and282. The cams 294 are coupled to the arms 286. The cams 294 can beconfigured to selectively open and close the compression members 272 and282 in accordance with conventional cam design methods.

In one alternative embodiment of the reaction unit, the compressionmembers 272 and 282 of each of the processing stations and insulatorsinclude holes 296 for receiving a cam 294 and a linear spring element298. Spring elements 298 each operate to bias a respective compressionmember in a direction away from the corresponding stationary member. Thecams 294, in combination with the springs 298, act to impartreciprocating motion to the actuators and regulate the compressingpressure on the sample vessel.

FIGS. 19A-19C illustrate a further embodiment of the reaction unit ofthe present invention. The reaction unit 350 includes nine openings 352for receiving up to nine sample vessels simultaneously. The reactionunit 350 includes three processing stations: a first processing station354, a second processing station 356, and a third processing station358. Thermal insulators 360A-360D are positioned adjacent each of theprocessing stations and at the top of the first processing station 354and the bottom of the third processing station. Top thermal insulator360A and bottom thermal insulator 360D are movable independent of thefirst processing station 354 and the third processing station 358,respectively. Intermediate thermal insulators 360B and 360C are coupledto the first processing station 354 and the second processing station356, respectively.

Each processing station comprises a stationary member 370 and acomplementary compression member 372 adapted to compress the samplevessel selectively within one or more of the openings 352 and therebymove the sample within the sample vessel. Each stationary member 370 hasa projection 374 aligned with one of the openings 352. The compressionmembers 372 are each provided with a projection 376, positioned on anopposite side of the opening 352. When a compression member 372 is slidon the corresponding stationary member 370, the projections 374 and 376engage and close the openings 352 therebetween.

Each compression member 372, as well as intermediate thermal insulators360B and 360C, include an arm 380 coupled by a cam 384 to a rotary shaft382. A stationary insulator member 362 is coupled, and aligned with anedge of each opening 352 on each stationary member 370. Each stationaryinsulator member 362 is inserted in each of the openings of a movableinsulator compression member 360 to react to compression and open orclose the opening. The shaft 382 is rotated by a stepper motor or aservo motor 386. The cams 384 translate the rotation of the shaft 382into linear reciprocal motion, which is imparted to the arms 380 toeffect selective opening and closing of the openings 352 and compressionof the sample vessels therein.

Each arm 380 includes an inner shaft 390 received within an outer sleeve392. A spring 394 is interposed between the inner shaft 390 and therespective compression member or thermal insulator. A second spring 396is positioned on an opposite side of the respective compression memberor thermal insulator. The spring 394 cooperates with the second spring396 to allow the compression member or thermal insulator to “float”along the axis of the arm 380. In this manner, the arm 380 can applysufficient force to the compression member or thermal insulator tocompress the sample vessel within an opening 352 and, thereby, displacesubstantially all of the sample from the compressed portion of thesample vessel. An increase of pressure within the sample vessel, forexample, from the compression of an adjacent portion of the samplevessel, however, can cause the sample to displace within the samplevessel through the compressed portion of the sample vessel, as thesprings 394 and 396 will allow small axial movements of the compressionmember or thermal insulator.

Each stationary member 370 and compression member 372 can be providedwith an embedded thermal energy transfer device 398 for each opening 352to apply thermal energy to the sample vessel within the opening 352. Inaddition, the stationary member 370 and compression member 372 caninclude temperature sensors 399 associated with each energy transferdevice 398 to monitor the temperature of the sample vessel.

FIGS. 15A and 15B illustrate embodiments of a sample vessel 16 accordingto the present invention. The illustrated sample vessel 16 is a closedtubule system that provides a disposable, single use container andreaction vessel for the sample. The sample vessel 16 can be constructedof a resiliently compressible, flexible, and ultra-high strengthmaterial, such as polyethylene or polyurethane. The sample vessel 16 canhave a seamless, flattenable cross-sectional profile and thin-walledconstruction that is optimized for fast and uniform heat transfer, formaximum surface contact with the sample, and for high pressureresistance. Preferably, the walls are constructed to converge when thesample vessel is compressed in a direction perpendicular to thelongitudinal axis of the sample vessel such that the volume of thecompressed portion of the sample vessel decreases and the ratio of thesurface area to the volume of the compressed portion increases, withoutfracturing of the sample vessel. In one illustrative preferred practice,the walls of the sample vessel 16 have a wall thickness of approximately0.01 mm to 0.5 mm. Experimental results indicate that constructing asample vessel having a wall thickness within this preferred rangesignificantly increases the efficiency of heat transfer to the sample.In an alternative embodiment, a two-layer wall structure can be used,with the inner layer providing bio-compatibility, using material such aspolyethylene or polyurethane, and the outer layer providing lowerpermeability, using material such as high density polyethylene oraluminum foil. In addition, the material selected to construct theportions of the wall of the sample vessel, such as a detection segmentof the sample vessel 16, can be optically transmissive over a selectedwavelength range to facilitate optical analysis of the sample within thesample vessel.

The sample vessel 16 can be divided into multiple segments by one ormore pressure gates 32. In the case of PCR testing, for example, thesample vessel can be divided into a sample collection segment 205, asample pretreatment segment 206, a sample reaction segment 208, and asample analysis segment 210. The illustrated pressure gates 32 areinternal to the tubule structure of the vessel 16 and provide a fluidtight seal between the segments of the sample vessel 16, under normaloperating conditions. Preferably, the pressure gates 32 open upon theapplication of pressure greater than a certain value, for example,approximately 3 atmospheres. When external pressure is provided to onesegment, the pressure gate 32 can open, allowing the sample to flow fromthe high pressure compartment to the low pressure compartment.

The sample vessel 16 can include a handling portion having a generallyrigid construction to facilitate handling of the sample vessel. Thehandling portion can be coupled to one or more of the segments of thesample vessels used to contain the sample. For example, the handlingportion can be a cylindrical sleeve constructed of a generally rigidmaterial, such as a plastic or a metal, that is sized and shaped to fitover one or more of the segment of the sample vessel. In one embodiment,the cylindrical sleeve can be removable and replaceable. Alternatively,the handling portion can be a rigid segment, such as a rigid ring,positioned at an end of the sample vessel or between two segments of thesample vessel. In the embodiments illustrated FIGS. 15A and 15B, thehandling portion is a segment of the sample vessel having an increasedwall thickness. For example, the sample collection segment 205 and thesample pretreatment segment 206 have a wall thickness greater than thewall thickness of the reaction segment 208. The increased wall thicknessprovides sufficient rigidity to the sample collection segment 205 andthe sample pretreatment segment 206 to facilitate handling of the samplevessel 16. In one embodiment, the wall thickness of the handling portionis greater than 0.3 mm.

The sample vessel 16 can include an instrument, such as a samplingpipette or a needle 107, for direct collection of the sample to betreated and analyzed within the sample vessel 16, as illustrated in FIG.15A. The needle 207 can be positioned at one end of the sample vessel 16and can be connected to the sample collection chamber 205 through aconduit 209 formed in the wall of the sample vessel 16. A needle cover211 can be provided to secure the needle 207 prior to and after use. Theneedle cover 211 can be, for example, a penetrable rubber cover or aremovable plastic cover.

In another embodiment, illustrated in FIG. 15B, a sampling instrument214, such as a pipette, a stick, or a tweezer, can be coupled to a cover212 that selectively closes the conduit or opening 209 formed in thewall of the sample vessel. The cover 212 can include a reservoir 216 forcontaining a reagent and a sample during sampling. For sampling, thecover 212 can be removed from the sample vessel to expose the samplinginstrument 214. The sampling instrument 214 can be used to collect thesample, by pipetting, swabbing, or gathering the sample, for example,and then the sampling instrument 214 can be inserted into the samplecollection segment 205 through the conduit 209. The sample can then beintroduced to the sample collection segment 205 by compressing the cover216 to displace the sample from the reservoir 216. Alternatively, thesample can be introduced to the sample collection segment 205 or toanother segment of the sample vessel, depending of the segments presentin the sample vessel, after collection by a separate instrument.

Sample vessel 16 can be particularly suited for PCR testing using thesample processing device of the present invention, as described above.For example, nucleic acid extraction can be performed within the samplepretreatment segment 206 of such a sample vessel 16. A cell lysesreagent, for example, GENERELEASER® from Bioventures, Release-IT™ fromCPG Biotech, or Lyse-N-Go™ from Pierce, or other extraction reagents canbe introduced to the pretreatment segment 206 to extract nucleic acidfrom the initial sample. Extraction reagents can be stored within thepretreatment segment 206 or can be delivered to the segment.Additionally, one or more filters can be positioned within thepretreatment segment 206 of the sample vessel to separate nucleic acidfrom unwanted cellular debris.

After incubation of the sample for certain time period, a portion ofpretreated sample can be moved into the reaction segment 208. For areaction sample volume of approximately 5 μl-25 μl, a PCR reactionsegment 208 of the sample vessel 16 according to one illustrativepractice of the invention has a wall thickness, indicated by referencecharacter t in FIG. 15A, of approximately 0.01 mm-0.3 mm, a diameter ofless than approximately 6 mm, and a length of less than approximately 30mm. PCR reagents, such as nucleotides, oligonucleotides, primers andenzymes, can be pre-packaged in the reaction segment or reactionsegments 206, or can be delivered, for example, through the walls of thesample vessel using a needle, using for example, a reagent injectorcartridge described below, before moving the sample into the segment.

A pre-packaged reagent storage segment 214 can be used to stored apre-packaged reagent. Such a reagent storage segment can be formedbetween any two adjacent processing segments and may store any reagentneeded for a reaction. For example, the reagent storage section 214 canstore PCR reagents, while reagent storage sections 236 and 244,described below, may include detection reagents. If the reagent storagesegment 214 is utilized, the sample vessel 16 can be compressed at thereagent storage segment 214 to displace the reagent into thepretreatment segment 206. Alternatively, the sample can be moved fromthe pretreatment segment 206, through the reagent storage segment 214where mixing with the reagent, to the reaction segment 208.

A self-sealing injection channel 218 can be formed in the sample vesselto facilitate delivery of reagent or other materials to the samplevessel, as illustrated in FIG. 16. The illustrated self sealinginjection channel 218 is normally substantially free of fluidic materialand is capable of fluid communication with the adjacent segment in thevessel. An injection of reagent through an injection channel occurspreferably prior to moving any sample into the segment to avoidcontamination. In addition, the sample treatment devices of theinvention can utilize a reaction cartridge 220 with a single or multipleneedles 222 in fluid communication with one or more reservoirs, asillustrated in FIG. 17. The reaction cartridge 220 can be used to injector deposit reagent or other materials, simultaneously, or sequentiallyinto multiple segments of the sample vessel. Suitable self-sealinginjection channels and reagent cartridges are described in U.S. Pat. No.6,318,191, incorporated herein by reference.

One skilled in the art will appreciate that while it may be preferablefor the wall of the sample vessel to be uniform along the circumferenceand the longitudinal axis of the vessel, only a portion of the wallalong the circumference and/or the longitudinal axis of the vessel needbe resilient and compressible and have the preferred thickness to affectflattening of the sample vessel. Thus, the sample vessel need not have auniform axial or circumferential cross-section.

PCR thermal cycling can be performed in the reaction segment 208 of thesample vessel 16. The thin walled, compressible construction of thesample vessel 16 greatly improves the rate and efficiency of thermalcycling. The construction of the sample vessel allows the vessel todeform or flatten readily, increasing thermal contact with the reactionunit of the device 10 and increasing surface/volume ratio of the samplewithin the sample vessel. As a result, the reaction mixture ramping rateis increased and thermal energy is more uniformly transferred to thesample.

PCR analysis can be performed in the sample vessel 16. For example,real-time detection methods can be used within the reaction segment 208;gel electrophoresis or other nucleic acid detection methods can be usedwithin the analysis segment 210 to analyze the sample. In the case ofgel electrophoresis, a gel can be introduced to the analysis segment 210to facilitate gel electrophoresis, as described above in connection withFIG. 14.

In one preferred embodiment, illustrated in FIG. 15A, the analysissegment 210 is divided into two electrophoresis capillaries, namely, asample capillary 230 and a control capillary 232, by adiametrically-central divider 234. Pressure gates 32 at either end ofthe capillaries control the movement of the sample and the reagents intoboth capillaries. Each capillary is filled with an electrophoresis gelsuch that gel electrophoresis can be performed simultaneously in bothcapillaries. A pair of electrodes 240, for both capillary 230 and 232,can be positioned within the walls of the sample vessel. A reagentstorage segment 236 can be provided at the proximal end of the samplecapillary 230 for storing reagent within the sample vessel prior to thesample entering the sample capillary 230. A control material can bestored in a control storage segment 242 positioned at the proximal endof the control capillary 232. A reagent can be stored in a reagentsegment 244 positioned at the distal end of the capillaries and incommunication with both the sample capillary 230 and the controlcapillary 232 for detection or display signal. The presence of thecontrol capillary 232 facilitates detection and analysis of the sampleby providing a basis of comparison for the sample analysis.

One skilled in the art will appreciate that the number of segmentswithin the sample vessel is dependent upon the sample being processedand the processing methods being employed. For example, in the case ofPCR testing, the number of segments within the sample vessel can bethree or more. Alternatively, thermal cycling and analysis may beperformed in one segment, reducing the number of segments to two. Incertain cases, an isothermal nucleic acid amplification method, forexample, only one segment may be necessary.

FIG. 18 illustrates a sample vessel 416 particularly suited for use in amulti-opening sample processing device such as, for example, the sampleprocessing device illustrated in FIG. 6. The sample vessel 416 includesan opening 420 for receiving the sample, a cap or closure 424 forselectively closing and sealing the opening 420, and a sample containingportion 426 within which the sample can be treated. The opening 420 isformed in a handling portion 428 that is preferably constructed of agenerally rigid or semi-rigid material, such as plastic or metal, tofacilitate handling of the sample vessel 416. The handling portion 428includes a collar 430 against which the cap 424 seats. Sample materialcan be introduced into the sample containing portion 426 of the samplevessel 416 through the opening 420. The collar 428 preferable tapersfrom a larger diameter to the smaller diameter of the sample containingportion 426. The sample containing portion 426 is preferable constructedof a resiliently compressible, flexible, and ultra-high strengthmaterial, such as polyethylene or polyurethane. The sample containingportion 426 can have a seamless, flattenable cross-section profile andthin-walled construction that is optimized for fast and uniform heattransfer, for maximum surface contact with the sample, and for highpressure resistance. In accordance with one embodiment, the samplecontaining portion 426 has a wall thickness of approximately 0.01 mm-0.3mm. Preferably, the sample containing portion 426 of the sample vessel416 is in a flattened state prior to introduction of the sample.Introduction of the sample to the sample containing portion 426 willcause the walls of the sample containing portion to separate and thevolume of the sample containing portion to increase. Compression of aselected portion of the sample containing portion 426 can cause thesample to displace to another portion within the sample containingportion along the length of the sample vessel. The surface of the samplevessel can be chemically treated to reduce a surface effect on thereaction.

The embodiments of the sample vessel described herein in connection withFIGS. 14-16 and 18, are not limited to use with the embodiments of thesample processing device described herein. The sample vessel of thepresent invention may be used with any sample testing or processingsystem. Likewise, the sample processing device of the present inventionis not limited to use with the sample vessels described herein. Othersample vessels may be used without departing from the scope of thepresent invention.

Certain changes may be made in the above constructions without departingfrom the scope of the invention. It is intended that all mattercontained in the above description or shown in the accompanying drawingsbe interpreted as illustrative and not in a limiting sense.

The present disclosure is also directed to sample vessels that may beutilized to collect and process one or more samples in a closed system.Exemplary samples that may be collected, processed, or otherwisecontained by the sample vessels disclosed herein include biologicalsamples, such as blood, urine, saliva, tissue, cell suspensions,microbial organisms, viruses, nucleic acids, and oligonucleotidessamples; soil; water; and any other sample materials that may be assayedusing known assays. The term “collection” as used herein generallyrefers to the extraction or gathering of the sample from a samplesource, the subsequent transfer of the sample into the sample vessel, orthe combination of extraction and subsequent transferring of the sampleinto the sample vessel. Exemplary sample gathering may includepipetting, biopsying, swabbing, drawing a fluid sample or other methodsfor extracting a sample from a sample source. Exemplary sample sourcesmay include humans or other animals, plants, water sources, cellcultures, food, other sample vessels, and chemical and biologicalassays. Sample sources may also include interim storage media, forexample, test tubes, syringes, absorbent applicators and other interimstorage media for containing a sample of interest. The term “processing”as used herein generally refers to the preparation, treatment, analysis,and/or the performance of other testing protocols or assays on a contentof the sample vessel in one or more steps. Exemplary processing stepsinclude, for example: displacing a content, e.g., the sample or areagent, of the sample vessel within the sample vessel to, for example,adjust the volume of the content, separate content components, mixcontents within the sample vessel; effecting a chemical or biologicalreaction within a segment of the sample vessel by, for example,introducing a reagent to the sample, agitating the sample, transferringthermal energy to or from the sample, incubating the sample at aspecified temperature, amplifying components of the sample, extracting,separating and/or isolating components of the sample; or analyzing thesample to determine a characteristic of the sample, such as, forexample, the quantity, count, volume, mass, concentration, or expressionlevel of a molecule, a target, a content, a marker or an analyte,binding activity, nucleic acid sequence, or nucleic acid size or otheranalyte size, of the sample. One skilled in the art will appreciate thatthe forgoing exemplary processing steps are described herein forillustrative purposes only. Other processing steps may be employedwithout departing from the scope of the present disclosure.

FIGS. 20A-20C illustrate an exemplary embodiment of a sample vessel 1000for collecting and processing one or more samples. The illustratedsample vessel 1000 comprises a tubule 1200 that provides a disposable,single use container and collection and processing vessel for thesample. The tubule 1200 may be constructed from any biocompatiblematerial and may be manufactured by injection molding, insert molding,dip molding, blow-molding, extrusion, co-extrusion, lamination,assembling from a sheet material, or other processes generally used tomanufacture medical devices and implants. The tubule 1200 may receivesample in solid or liquid form and, in certain embodiments, may be sizedto collect and/or process sample volumes in the range of 2 microlitersto 2000 microliters.

The tubule 1200 may be used with any known sample testing or processingsystem, including, for example, the systems described in U.S. Pat. No.6,318,191, U.S. patent application Ser. No. 10/605,459, filed Sep. 30,2003, which is a continuation of U.S. patent application Ser. No.09/339,055, abandoned, and U.S. Pat. No. 6,780,617. Each of theaforementioned patents and patents applications is incorporated hereinby reference.

In the exemplary embodiment illustrated in FIGS. 20 and 21, the tubule1200 may include an opening 1400 for receiving a volume of samplematerial. The tubule 1200 may include a compressible segment 1600 havinga wall 1800 constructed at least partially from a material havingsufficient flexibility to permit compression of the opposed segments ofthe wall 1800 into contact. For example, the wall 1800 may beconstructed to converge when the compressible segment 1600 of the tubule1200 is compressed in a direction perpendicular to the longitudinal axisof the tubule such that the volume of the compressed segment 1600 of thetubule 1200 decreases, without fracturing of the sample vessel. Thewalls 1800 of the compressible segment 1600 may be constructed of aresiliently compressible, flexible, and ultra-high strength material,such as polyethylene, polyurethane, polyvinyl chloride, polypropylene,or any other plastic material suitable for biomedical or chemicalassaying applications. In one illustrative embodiment, the walls 1800 ofthe compressible segment 1600 have a wall thickness of approximately0.01 mm to 0.5 mm. Experimental results indicate that constructing acompressible segment of a tubule having a wall thickness within thisrange significantly increases the efficiency of sample processing, suchas heat transfer to the sample and sample transfer between the segments,and detection. In the illustrated embodiment, the compressible segment1600 of tubule 1200 extends the entire length of the tubule 1200.Alternatively, as discussed below, the tubule 1200 may include one ormore discrete compressible segment 1600 spaced apart from one or moresegments having different (e.g., non-flexible) properties.

In other exemplary embodiments, the tubule 1200 may comprise amulti-layer wall structure. For example, the tubule 1200 may include aninner layer providing bio-compatibility, using material such aspolyethylene or polyurethane, and an outer layer providing lowerpermeability, using material such as high density polyethylene or ametal foil, such as aluminum foil or a metal deposition. One skilled inthe art will appreciate that one or more additional layers may also beemployed, depending on, for example, the sample type, the reagent(s)employed, and the assay(s) being performed.

The material selected to construct portions of the wall of the tubule1200, for example an optional detection segment of the tubule 1200, canbe optically transmissive over a selected wavelength range to facilitateoptical analysis of the sample within the tubule 1200.

The sample vessel 1000 of the exemplary embodiment illustrated in FIGS.20A-20C may comprise a general rigid container 2000 for receiving all orat least a portion of the tubule 1200. In the illustrated embodiment,the container 2000 is sized to receive the complete length of the tubule1200. The container 2000 may be constructed of a material havingincreased rigidity compared to the material of the tubule 1200 tofacilitate handling of the tubule 1200. In certain embodiments, thecontainer 2000 may be constructed of a material having a lowerpermeability than the material of the tubule 1200. In the illustratedembodiment, the container 2000 is a glass vacuum tube. Suitable glassvacuum tubes are available under the trademark VACUTAINER® fromBecton-Dickenson. The sample vessel 1000 can be used in a manner similarto a glass vacuum tube to collect a sample, such as a blood sample. Acontainer 2000 may be optionally used with any of the tubule embodimentsdisclosed herein.

The sample vessel 1000 may comprise an interface 3000 that is in fluidcommunication with the opening 1400 in the tubule 1200. The interface3000 may permit collection of the sample within the tubule 1200 byfacilitating delivery of the sample material to the tubule 1200 throughthe opening 1400. In certain exemplary embodiments, the interface 3000may include an instrument for collecting the sample form a samplesource. In the exemplary embodiment illustrated in FIGS. 20A and 20B,the interface 3000 is a stopper 3200 that may be coupled to the tubule1200 and may selectively seal the opening 1400 in the tubule 1200 tofacilitate collection of the sample from a separate instrument. In theexemplary embodiment, the stopper 3200 is removably and replaceablyconnected to the rigid container 2000 and seals an opening 2200 in thecontainer 2000. The stopper 3200 may include a first annular portion3400 having an opening 3600 sized and shaped to receive the tubule 1200in a fluid tight relationship. The first annular portion 3400 is furthersized and shaped to engage the walls of the container in a fluid tightrelationship. The stopper 3200 may include a second annular portion 3800that has a diameter greater than the diameter of both the first annularportion 3400 and the container 2000. The opening 3600 extends throughthe second annular portion 38 to form an interface channel 3700. Apenetrable, self-sealing portion 4000, such as a self-sealing membrane,may be provided to selectively seal the opening 3600 and, thus, permitselective transfer of the sample (from, for example, the samplecollection instrument) through the interface channel 3700 into thetubule 1200. The self-sealing portion 4000 may be constructed of anybiocompatible, resilient, self-sealing material that can be penetratedby a needle or other sample collection instrument. Suitable materialsmay include rubber and silicon. In certain embodiments, the stopper 3200may be constructed completely from a biocompatible, resilient,self-sealing material such as rubber or an elastomeric polymer. Theinterface channel 3700 may taper or otherwise narrow through thecross-section of the stopper 3200 to provide a guide for a needle orother instrument transferring the sample to the tubule 1200.

Alternatively, the interface 3000 may include other mechanisms forselectively sealing the opening 1400 in the tubule 1200. For example,the interface may include a self-sealing elastomeric duckbill closure.Alternatively, the interface 3000 may include a valve for selectivelyclosing and opening the interface channel 3700.

The sample vessel 1000 may include a clamp 5000 for compressing thecompressible segment 1800 of the tubule to adjust the volume of thetubule 1200. The clamp 5000 may be configured to compress opposing wallportions of the compressible section 1600 into contact thereby dividingthe tubule 1200 into two segments, 1600A and 1600B, as best illustratedin FIG. 20B. When the clamp 5000 is employed, the segment 1600A remainsin fluid communication with the interface channel 3700 and segment 1600Bis sealed from segment 1600A by the clamp 5000. Once the sample isdelivered to the segment 1600A of the tubule 1200, the clamp 5000 may beremoved, providing additional volume in the tubule 1200 that may permitfuture segmentation of the tubule and displacement of the sample withinthe tubule 1200 by compression of the tubule 1200.

The clamp 5000 may be positioned at any location along the longitudinalaxis of the tubule 1200. Additional clamps may also be employed todivide the tubule into additional segments. In illustrated exemplaryembodiment, the clamp 5000 is disk-shaped and includes a radial slot5200 that is sized to receive the tubule 1800 in a compressed state. Oneskilled in the art will appreciate that other devices may used tocompress and, thereby, divide the tubule 1200.

In certain exemplary embodiments, the tubule 1200 may be wholly orpartially evacuated to place the lumen 4200 of the tubule 1200 undernegative pressure, e.g., at a pressure less than atmospheric pressure,to facilitate fluid flow into the tubule 1200. Negative pressure can begenerated by, for example, compressing the tubule 1200 to collapse thelumen 4200. An apparatus suitable for compressing the tubule isillustrated in FIGS. 36A-36C, described below. Alternatively the tubule1200 may be compressed by hand. The tubule 1200 may also be manufacturedto include a negative pressure.

In certain embodiments, the container 2000 may be wholly or partiallyevacuated to a negative pressure. For example, the container 2000 may beevacuated to inhibit loss of negative pressure within the tubule 1200and to hold the shape of the tubule 1200 during storage.

A reagent may be pre-packaged in the tubule 1200 or can be introduced tothe tubule 1200 after the sample is introduced to the tubule 1200. Forexample, a reagent can be introduced using a reagent injector cartridgeassociated with the sample processing system, by a needle, or by anotherdevice capable of fluid communication with the tubule 1200. The reagentcan be, for example, an anticoagulant, a cell lyses reagent, anucleotide, an enzyme, a DNA polymerase, a template DNA, anoligonucleotide, a primer, an antigen, an antibody, a dye, a marker, amolecular probe, a buffer, or a detection material. The reagent can bein liquid or solid form. In the case of a solid reagent, the reagent maybe coated onto the walls of the tubule 1200.

In certain exemplary embodiments, the interface 3000 may include one ormore chambers 44 that are in fluid communication with the tubule 1200 toselectively receive a volume of fluid, such as the sample material or areagent, from the tubule 1200. In certain exemplary embodiments, thechamber 4400 may be evacuated or constructed to have a substantiallysmall initial volume and may be expendable when receiving fluid. Thechamber 4400 can be used as a waste container to receive and storeoverflow sample, wash buffer, or reaction mixture during the sampleprocessing. For example, compressing a segment of the tubule 1200 maymove a portion of the sample to the chamber 4400.

In the exemplary embodiment illustrated in FIGS. 21A-21C, for example,the stopper 3200 includes an annular chamber 4400 that is in fluidcommunication with the interface channel 3700 in the stopper 3200, and,thus, the tubule 1200, through a pressure gate 4800. In certainembodiments described herein, one or more pressure gates may be employedto selectively control the flow of fluid between segments, lumens, andother portions of the tubule, as well as between the tubule and externaldevices. For example, the illustrated pressure gate 4800 provides afluid tight seal between the chamber 4400 and the interface channel 3700under normal operating conditions. The pressure gate 4800 may open uponthe application of a fluid pressure greater than a certain thresholdpressure, for example, approximately 3 atmospheres. When a fluidpressure equal to or greater than the threshold pressure is applied tothe pressure gate 4800, the pressure gate 4800 can open, allowing thesample or a reagent to flow from the high-pressure compartment, e.g.,from the tubule 1200 or from the chamber 4400, to the low-pressurecompartment. In certain embodiments, the pressure gate may bereversible, i.e., the pressure gate may be configured to re-close if thefluid pressure is reduced to value less than the threshold pressure. Inother embodiments, the pressure gate may be irreversible, i.e., thepressure gate may be initially closed and may remain open once opened.For example, once a threshold pressure is exceeded the irreversiblepressure gate remains open, even if the pressure applied to the pressuregate is reduced to below the threshold pressure. One example of anirreversible pressure gate is the pressure gate described below inconnection with FIGS. 22A-22B.

In the illustrated embodiment of FIGS. 21A and 21B, the pressure gate4800 is a slit formed in the stopper 3200 between the interface channel3700 and the chamber 4400. The material forming the stopper 3200 may beselected to be sufficiently flexible and resilient to allow the slit toopen at the threshold pressure and to close at pressures lower than thethreshold pressure.

A label 6000 identifying the sample within the sample vessel 1200 may beattached to the interface 3000, the container 2000, or the tubule 1200.The label 6000 can be a bar code or other indicia for identifying thesample.

FIGS. 22A and 22B illustrate another exemplary embodiment of a samplevessel 10000. The sample vessel 10000 comprises a tubule 11200, whichcan be analogous in construction to the tubule 1200, having a pluralityof lumens 14200A and 14200B. The plurality of lumens 14200A and 14200Bcan be separated by a pressure gate 148 that permits selective fluidflow between the lumens 14200A and 14200B. FIG. 22A illustrates thepressure gate 14800 in a closed position and FIG. 22B illustrates thepressure gate in an open position that permits fluid flow between thelumens.

In the exemplary embodiment, the lumens 14200A and 14200B are parallelto each other and extend in a direction generally parallel to thelongitudinal axis of the tubule 1200. One skilled in the art willappreciate that other lumen orientations are possible. The lumens 14200Aand 14200B may be uniform in size (e.g., diameter, width, length) andshape or, alternatively, the lumens 14200A and 14200B may be differentin size and shape, as in the illustrated embodiment. For example, in theillustrated embodiment, the lumen 14200B has a smaller diameter than thelumen 14200A. Although two lumens are illustrated in the exemplaryembodiment, one skilled in the art will appreciate that the tubule 1200may be constructed of any number of lumens.

The pressure gate 14800 in the present embodiment is coextensive withthe lumens 14200A and 14200B, i.e. the pressure gate 14800 extends alongthe entire length of the lumens. Alternatively, the pressure gate 14800may extend along only a portion or portions of the lumens, particularlyin embodiments in which the tubule 1200 is segmented into discretelongitudinally extending segments, as in the case of the embodimentillustrated in FIGS. 26A-26C. In such embodiments, one or more pressuregates may be provided between adjacent lumens.

In the exemplary embodiment, the opposed portions of the wall 11800 ofthe tubule 11200 are compressed into contact to form a longitudinallyextending seam 17000 that divides the tubule 11200 into two lumens,lumens 14200A and 14200B. In addition to dividing the tubule 11200 intomultiple lumens, the seam 17000 may further provide an irreversiblepressure gate, pressure gate 14800, between the lumens 14200A and14200B. The seam 17000 may be formed by mechanically clamping orotherwise compressing a cylindrical tubule or by applying vacuumpressure to the interior of a cylindrical tubule. Alternatively, theseam 17000 may be formed during manufacturing of the tubule by, forexample, extrusion, molding, or lamination processes. The opposed wallportions that are compressed into contact to form the seam 17000, andthe pressure gate 14800, may be bonded together by mechanical orchemical bonding, by heating sealing, for example, by bringing hotsurfaces into contact with the tubule wall immediately after extrusion,by ultrasonic welding, by mechanical interlocking, or both otherconnection mechanisms, to create the irreversible pressure gate 14800.

The pressure gate 14800 is initially in a closed configuration thatinhibits fluid flow between the lumens 14200A and 14200B. The pressuregate 14800 may open by separating the compressed opposed walls formingthe pressure gate 14800. Applying a threshold pressure to the pressuregate 14800, as described above, may open the pressure gate 14800.Alternatively, energy may be applied to the pressure gate 14800 toweaken the bond between the compressed opposed walls. For example,thermal energy or light, e.g., ultra-violet light, may be applied to thepressure gate 14800 or to selected portions or all of the tubule 11200.The threshold pressure and/or the amount energy to open the pressuregate 14800 may vary depending on the type and strength of the bond.Alternatively, the bond between the compressed opposed wall portions maybe weakened or broken by chemical reaction with reagent or the sample.

In certain exemplary embodiments, one or more of the lumens may includeone or more reagents. Reagents may be provided to one or more lumensprior to sample collection, e.g., one or more reagents pre-packaged withthe tubule, or after sample collection. In the exemplary embodimentillustrated in FIGS. 22A and 22B, for example, a reagent may be providedin lumen 14200B. Lumen 14200A may be utilized for sample collection andprocessing. Sample collection may occur with the pressure gate 14800 ina closed configuration, as illustrated in FIG. 22A. Upon transfer of thesample to lumen 14200A, the pressure gate 14800 may be openedautomatically due to release of pressure within the lumen 14200A, orselectively by applying energy to the pressure gate and/or a thresholdfluid pressure. In other embodiments, the lumen 14200A or 14200B may becompress to provide the threshold pressure. Upon opening the pressuregate 14800, the reagent(s) can mix with and interact with the sample inthe lumen 14200A, as illustrated in FIG. 22B. Automatic release of thepressure gate 14800 and mixing of the reagent with the sample may bebeneficial in certain applications, such as the mixing of ananticoagulant with a blood sample.

FIG. 23 illustrates another embodiment of a multi-lumen tubule 11200that includes three lumens, namely a first lumen 14200A, a second lumen14200B, and a third lumen 14200C. Each lumen may be separated a pressuregate 14800, for example, an irreversible pressure gate, as describedabove. Each of the lumens 14200A, 14200B, and 14200C may be providedwith one or more reagents and/or may be used for sample collection andprocessing. For example, second lumen 14200B may be provided with one ormore prepackaged reagents and first lumen 14200A may be used for samplecollection and processing. Upon sample collection in first lumen 14200A,pressure gate 14800A may be opened allowing fluid communication betweenthe second lumen 14200B and the first lumen 14200B. FIG. 23 illustratesthe pressure gate 14800A in an open configuration. Lumen 14200C may beutilized as an injection channel for receiving one or more reagents,typically, but not necessarily, after sample collection in first lumen14200A. The lumen 14200C may be free of sample material until pressuregate 14800A is transitioned to an open configuration. FIG. 23illustrates pressure gate 14800B in a closed configuration that inhibitsfluid communication between third lumen 14200C and first lumen 14200A.Reagent may be delivered to the third lumen 14200C by a needle 19000,such as a needle from a reagent injection cartridge, or by otherinstruments that can penetrate the lumen or otherwise provide fluidcommunication between a reagent source and the lumen 14200C. The lumen14200C may be free of sample and reagent material until reagent isinjected to avoid cross contamination of the injection needle 19000. Theportion of the wall 11800C proximal the third lumen 14200C may beconstructed of a resilient, self-sealing material to facilitatere-sealing of the wall 11800 after penetration to deliver reagent.

One or more lumens of the tubule 11200 may include a reinforced wallportion 17100, as illustrated in FIG. 24. The reinforced wall portion17100 may have an increased wall thickness compared with the remainderof the tubule wall 11800 to facilitate needle penetration andre-sealing. For example, the reinforced portion may have a wallthickness of approximately 1 mm to 5 mm greater than other portions ofthe wall. The reinforced portion 17100 may be constructed from adifferent material, having increased strength and/or resiliency, forexample, than the remainder of the tubule wall 11800. Needle guides17200 may be provided to direct needle penetration and inhibit tearingof the tubule wall 11800.

FIGS. 25A-25E illustrate another exemplary embodiment of a multi-lumentubule 11200 that includes a pair of parallel lumens, namely first lumen14200A and 14200B. In the illustrated embodiment, the lumens 14200A and14200B are connected parallely by a thin layer fluid channel 17600 inthe form of a slit opening that extends the length of the tubule 11200.Although one fluid channel is illustrated, additional fluid channels maybe provided. The fluid channel 17600 permits the sample to be movedbetween the first lumen 14200A and the second lumen 14200B and to occupyboth lumens simultaneously. For example, during sample collection,portions of the sample, or the entire sample, can be transferred fromthe opening 11400 along the length of the first lumen 14200A, throughthe fluid channel 17600, and along the length of the second lumen14200B. FIGS. 25B-25E illustrate the flow of a sample, in fluid form,through the first lumen to the end of the first lumen due to relativelylow flow resistance of the lumen relative to the fluid channel 17600(FIG. 25B), through a portion 17400 of the fluid channel 17600 distal tothe opening in the first lumen 14200A (FIG. 25C), along the fluidchannel 17600 and through the second lumen 14200B (FIG. 25D) to fillboth lumens (FIG. 25D). In embodiments in which a solid reagent ispacked into the lumens 14200A and/or 14200B of the tubule 11200, flow ofthe sample through the lumens via the fluid channel 17600 can facilitatemixing of the solid reagent with the sample. For example, in the case ofblood samples, the inventors have determined that by allowing the bloodsample to flow through the first lumen 14200A and the second lumen14200B via the fluid channel 17600 can improve mixing of the sample withan anticoagulant coated on the inner walls of the two lumens.

FIGS. 26A-26C illustrate another exemplary embodiment of a multi-lumentubule 11200 having three parallel lumens, namely a first lumen 14200A,a second lumen 14200B, and a third lumen 14200C. In the exemplaryembodiment, each lumen of the tubule 11200 is divided into a pluralityof longitudinally extending segments 18000. For example, the third lumen14200C, illustrated in cross-section in FIG. 26B, includes five segments18000A-E. Each of the segments 18000 can be used for one or more samplecollection and/or sample processing steps, including the processingsteps described above. In PCR (polymer chain reaction) testing, forexample, one segment may used for sample collection, one segment may beused for sample pretreatment, e.g., nucleic acid extraction, one or moresegments may used for sample processing, e.g., thermocycling, and one ormore segments may be used for sample analysis. Any number of segmentsmay be provided. In addition, one or more segments may be used to storereagent or as an injection channel for the delivery of reagent. Thenumber of segments may be varied depending of the sample being processedand the processing steps selected.

Each of the segments 18000 may be separated by a seal 18200 thatprovides a temporary or permanent fluid seal between adjacent segments18000. A seal 18200 may be a pressure gate, such as the reversible andirreversible pressure gates described above. Alternatively, a seal 18200may be formed by bonding or fusing of compressed opposed wall sectionsof the tubule. The seal 18200 may be formed by applying energy, such asthermal energy or RF energy, by ultrasonic welding, or by using abonding agent. A clamp may also be applied to the exterior of the tubuleto compress the wall of the tubule and form a seal separating thesegments in the tubule. For example, the clamp may be anelectro-mechanical clamping mechanism as described below in connectionwith FIG. 29. Any other mechanism for provided an external compressiveforce on the tubule may be employed as the clamping mechanism. One ormore clamps may be provided as part of the sample processing system usedto process the sample within the tubule 11200. The segments may beconnected by one or more micro-fluidic flow channels that provideselective fluid connection between the segments, as described below. Aseal 18200 may be a filter disposed within the tubule to separateselected components of a fluid within the tubule from other segment orcomponents of the fluid within the tubule.

In the illustrated exemplary embodiment, the interface 3000 forfacilitating delivery of the sample to the tubule 11200 includes aneedle 18400 for direct collection of the sample to be processed withthe sample vessel 10000. The needle 18400 is positioned a proximal endof the tubule and is fluid communication with an opening in the tubule11200. In the illustrated exemplary embodiment, the needle 18400 is influid communication with an opening in the first lumen 14200A, however,the needle 18400 may be connected to any one or all of the lumens 14200of the tubule 11200. A removable and replaceable needle cover 18600 maybe provided to secure the needle 18400 prior to and after use.Alternatively, the needle cover 18600 may be connected by a hinge, asshown in FIG. 27, or by another mechanism that allows to the cover 18600to be moved into and out of position about the needle 18400. A needlesafety mechanism may be coupled to the needle and the sample vessel.

FIG. 28 illustrates another embodiment of the cover 18600 in which thesample collection instrument, e.g., the needle 18400, is connected tothe cover 18600. In the illustrated exemplary embodiment, the proximalend 18400A of the needle 18400 may be used for sample collection from asample source and the distal end 18400B of the needle 18400 may be usedto provide a fluid connection with an opening in the tubule 11200through interface 3000. For example, the distal end 18400B may be usedto penetrate a self-sealing membrane 4000 provided in the interface3000. In another embodiment, a cover 19000 may include a sampleinstrument in the form of a needle 18400 and may have a compressibleportion in fluid communication with the needle to facilitate drawing afluid sample into the needle 18400 and transferring the sample to thesample vessel 11000. Cover 19000 may be particularly useful as a fingerprick for collection a blood sample.

FIG. 29 illustrates a processing station 30000 of an exemplary sampleprocessing device, such as a sample processing device described in U.S.Pat. No. 6,318,191 and U.S. patent application Ser. No. 09/782,732,filed Feb. 13, 2001. The exemplary processing station 30000 includesmultiple compression members, namely first compression member 30200A,second compression member 30200B, and third compression member 30200C.Each compression member 30200 is adapted to compress a sample vessel,for example, the tubule 1200 of sample vessel 1000 described above, andthereby displace the contents of the sample vessel, e.g. reagent orsample, within the sample vessel. Although the exemplary processingstation 30000 is illustrated in connection with sample vessel 1000, oneskilled in the art will appreciate that any of the sample vesselsdisclosed herein may be used in with the exemplary processing station30000. A plurality of compression members 30200 may be oriented parallelto the longitudinal axis of the tubule 1200, as illustrated in FIG. 29A.Alternatively, a plurality of compression members 30200 may be orientedtransverse to the longitudinal axis of the tubule, i.e., orientedlatitudinally, as illustrated in FIG. 34B described below, or in otherorientations depending on the compression configuration desired. Adriver may be coupled to one or more of the compression members 30200 toselectively move the compression member into contact with the samplevessel. The driver can be, for example, an electromagnetic actuatingmechanism, a motor, a solenoid, or any other device for imparting motionon the compression members 30200. A stationary member 30400 or anothercompression member may be provided opposite compression member 30200.

A compression member 30200 may be employed to compress a portion of thewall 1800 of the tubule 1200 into contact with another portion of thewall 11800 of the tubule 1200 to form a seal in the tubule 1200 andthereby divide the tubule 1200 into multiple segments. In alternativeembodiments, a compression member 30200 may compress a portion of thewall 1800 of the tubule 1200 into proximity with another portion of thewall 1800 of the tubule 1200 to form a micro-fluidic channel 30600between segments of the tubule 1200. For example, in the embodimentillustrated in FIG. 29, compression member 30200B compresses a portionof wall 1800 into proximity with another portion of the wall to create amicro-fluidic channel 30600 that connects a first segment 18000A and asecond segment 18000B of the tubule 1200. The width of the micro-fluidchannel 30600 may be adjusted by displacing the compression member30200B towards or away from the tubule 1200. Micro-fluid channels may beformed having a gap less than 200 microns, preferably 10 to 30 microns.

The compression members 30200 may be arranged in a variety oforientations to compress the tubule 1200 into a variety ofconfigurations. For example, in FIG. 29B, the width of the micro-fluidicchannel 30600 extends across the entire width of the tubule 1200. Such acompressed configuration may be formed by a compression member 30200Bhaving a planar compression surface 30800 for engaging the tubule 1200that is sized to engage the entire compressed wall surface of thetubule. In other embodiments, the size or shape of the compressionsurface 30800 may be varied and the number and orientation ofcompression members 30200B may be varied. For example, FIG. 30Aillustrates a compressed tubule 1200 having a centrally located flowchannel 30600 that may be formed by a compression member 30200 having agroove formed on the bottom surface thereof or by three compressionmembers 30200 aligned transverse to the longitudinal axis of the tubule1200. A centrally positioned compression member may compress wallportion 1800A into proximity with an opposed wall portion, while a pairof compression members, one on either side of the central compressionmember, may compress side wall portions 1800B and 1800C, respectively.FIG. 30B illustrates a compressed tubule 1200 having a centrally locatedlumen 30600 formed by compressing the tubule 1200 into a non-planarconfiguration. In this illustrated embodiment, a triangular profile flowchannel is formed, which inherently forces a cell or particle to flowthrough the central line of the channel, thus reducing the need toregulate the tolerance in forming the flow channel. FIG. 30C illustratesa compressed tubule 1200 having a flow channel 30600 formed off-set fromthe center of the tubule 1200. In the illustrated embodiment, the flowchannel is formed on a lateral edge of the tubule 1200.

At least a portion of the wall of the tubule 1200 may be opticallytransparent to allow monitoring or detection of the sample or reaction.The transparent portion of the wall may be located in the flow channelsection, thus allowing the monitoring of sample or reaction under flowor through a thin layer of liquid, for processes such as counting cells,reaction hybridization, or detection, for example, microarray spots.

One skilled in the art will appreciate that while it may be desirable incertain applications for the wall of the tubules disclosed herein to beuniform along the circumference and the longitudinal axis of the tubule,only a portion of the wall along the circumference and/or longitudinalaxis of the tubule need be resilient and compressible. Thus, the tubuleneed not have a uniform cross-section, either along the longitudinalaxis or transverse to the longitudinal axis. In certain exemplaryembodiments, for example, a section of the wall of the tubule may beformed of a material selected to provide a property distinct from aproperty of another section of the wall. Exemplary properties that maybe varied include permeability, flexibility, hardness, resiliency,optical transparency, biocompatibility, surface smoothness of the innerwall, and surface chemistry of the inner wall, for example thehydrophobic or hydrophilic properties of the inner wall surface. Surfaceproperties may be rendered by coating with a layer of material, such asa thermoset urethane aired by UV energy or other cross linking methods.

FIGS. 34A and 34B illustrate an exemplary embodiment of a sample vessel1000 in the form of a tubule 1200 having wall sections 1800A that areformed of a material selected to provide a property distinct from aproperty of a plurality of other wall sections 1800B of the tubule 1200.Wall sections 1800A may be opposed to one another, as illustrated, orpositioned at other positions in the cross section of the tubule 1200.Wall sections 1800A may similar in size, shape and material properties,as illustrated, or may vary in size, shape, and material properties fromone another. In the illustrated embodiment, wall sections 1800A areselected from a material having sufficient flexibility to permitcompression of the tubule 1200, as illustrated in FIG. 34B. Wallsections 1800B are formed of a material having increased rigiditycompared to the material of wall sections 1800A. In the illustratedembodiment, wall sections 1800B preferably have sufficient rigidity toresist flexing during compression and thereby maintain a planarconfiguration. Wall sections 1800B may be opposed to one another, asillustrated, or positioned at other positions in the cross section ofthe tubule 1200. Wall sections 1800B may be similar in size, shape andmaterial properties, as illustrated, or may vary in size, shape, andmaterial properties from one another. In the illustrated embodiment, thewall sections 1800A and 1800B are spaced latitudinally, i.e., about thecircumference of the tubule 1200 and transverse to the longitudinalaxis. Wall sections 1800A and 1800B are interposed between one anotherin an alternating arrangement about the circumference of the tubule1200. Wall sections 1800A and 1800B may be formed from the same materialor a different material. For example, wall sections 1800A may be formedof a relatively low durometer polyurethane, for example, in the range offrom 40 A to 90 A depending on thickness, and wall sections 1800B may beformed of a polyurethane having a relatively higher durometer, forexample, in the range of from 40 D to 90D depending on thickness. Atubule having wall sections of varying properties may be manufactured byconventional extrusion, co-extrusion, injection molding, insert molding,dip molding, blow molding, transfer molding, or lamination processes.

During compression of the tubule 1200 illustrated in FIGS. 34A and 34B,the wall sections 1800A flex allowing a first wall section 1800B to bemoved into proximity or contact with second wall section 1800B′. Wallsections 1800B may provide improved sealing surfaces due to theincreased rigidity compared with wall sections 1800A. In addition, wallssections 1800B permit the formation of a precisely defined micro-fluidflow channel 30600, as illustrated in FIG. 34B. The increased rigidityof the wall sections 1800B allows for the formation of a smaller andmore uniform flow channel than more flexible wall sections. FIG. 34Billustrates the formation of a micro-fluidic flow channel 30600 betweensegments 18000A and 18000B of the tubule 1200. In the illustratedembodiment, the compression members 30200A-C are oriented transverse tothe longitudinal axis of the tubule to form a flow channel 30600 thatextends latitudinally, i.e., transverse to the longitudinal axis,between first segment 18000A and second segment 18000B.

In other exemplary embodiments, the number of wall sections of differingproperties may be varied. For example, a single wall section 1800Bhaving increased rigidity may be provided or three or more wall sectionshaving increased rigidity may be provided.

In certain exemplary embodiments, a flow channel 30600 may be pre-formedin a section of the wall 1800 of the tubule as illustrated in FIGS. 31and 32A-B. The pre-formed flow channel 30600 may be a groove 31600formed in a wall section of the tubule 1200. The groove 31600 may beformed by scoring or etching the wall 1800 of the tubule 1200 or may beformed during the extrusion or molding of the tubule 1200. The groove31600 in the illustrated embodiments extends longitudinally, however,the groove 31600 may be formed in any direction, includinglatitudinally. More than one groove 31600 may be provided. The groove31600 may have a variety of cross-section shapes and sizes. In theembodiment illustrated in FIG. 31, the groove 31600 has a triangularcross-section. In the embodiment illustrated in FIGS. 32A-32B, thegroove 31006 has a rectangular cross-section. The cross-sectional sizeof the groove 31600 can be selected based on desired shear rate profileof the flow channel 30600.

The groove 31600 may be formed in any section of the wall 1800 of thetubule 1200. For example, the groove 31600 may be formed in a wallsection 1800B having increased rigidity compared to other wall sectionsof the tubule 1200, as is the case for the illustrated embodiments ofFIGS. 31 and 32A-B. During compression of the tubule 1200, asillustrated in FIG. 32B, the wall section 1800B contacts wall section1800B′ to provide a fluid tight seal. Groove 31600 provides a flowchannel 30600 that extends longitudinally through the fluid tight seal.

FIGS. 33A and 33B illustrate an exemplary embodiment of a sample vessel40000 comprising a tubule 41200 having a plurality of flow channels30600 and one or a plurality of depressions 40800 formed on an interiorwall surface 41000 of the wall 41800 of the tubule 41200. Eachdepression 40800 can form a micro-cup during compression of the tubule41200 that can hold a fixed volume of sample or reagent. The volume of adepression forming a micro-cup can be from 0.1 microliter to 10microliter, preferably, from 0.5 microliter to 4 microliter. A patternof one or more grooves 31600 and depressions 40800 may be formed on theinterior wall surface 41000 of the tubule 41200 and may interconnect toprovide a network of micro-cups interconnected by micro-fluidic flowchannels 30600, as best illustrated in FIG. 33B. Such a network may beused to perform a variety of processing steps within one or moremicro-cups and may permit the transport of small, precise volumes ofsample and reagent between the micro-cups via micro-fluid flow channelsby selectively compressing the tubule 41200. The network of grooves anddepressions may be formed using semi-conductor processing techniques.For example, a mask pattern may be applied to an interior wall surfaceof the tubule 1200 using conventional photolithographic techniques. Thegrooves and depressions may then be formed by etching or otherwiseremoving portions of the interior wall surface based on the patternimaged onto the interior wall surface. It may be desirable to form thenetwork of grooves and depressions on a planar substrate 41800Aconstructed of a material suitable for use in the tubule 41200, asillustrated in FIGS. 33A and 33B. A second layer 41800B of material canbe attached to the planar substrate 41800A to form the wall 41800 of thetubule 41200.

Referring to FIGS. 33A and 33B, one or more sample or reagent processingdevices 41400 may be provided on the interior wall surface 41000 of thetubule 41200. For example, a microarray device may be embedded on theinterior wall surface 41000 of the tubule 41200. An exemplary microarraydevice 41400 may comprise a plurality of reagent coated zones forsimultaneous analysis of a plurality of analytes within a sample. Theprocessing device 41400 may also be a micro-fluid device or alab-on-a-chip device, or any other device for processing a sample. Theprocessing device 41400 may be interconnected with one or moredepressions 40800 or other processing devices via flow channels 30600.Any number of processing devices of any type may be provided in thetubule 41200.

Referring to FIG. 37, a sample vessel 70000 comprising a tubule 71200divided into multiple segments 78000A-C. Segment 78000B may beconstructed of a rigid, generally non-flexible material and may have aprocessing device, such as a microarray 71400, embedded on the interiorwall thereof. The segment 78000B may provide a pre-formed flow channelbetween two compressible segments 78000A and 78000C. By alternatelycompressing the two flexible segments 70800A and 78000C, the sample mayflow through the flow channel 70600 to facilitate high efficienthybridization or binding of analytes to the reagent spots of themicroarray 71400. A flow channel 706 having a small gap may alsoincrease wash efficiency as a laminar flow is formed.

FIGS. 35A-35E illustrates an exemplary embodiment of a sample vessel1000 comprising a tubule 1200 and an adapter 50000 that is connected tothe tubule 1200 of the sample vessel 1000. The adapter 50000 may beprovided to facilitate handling of the sample vessel 1000 and/or tofacilitate connection of the tubule 1200 to an external device, such asa micro-fluid device, a lab-on-a-chip device, a microarray device, areagent source, another sample vessel, or any other device suitable forcontaining or processing a sample. In the illustrated embodiments, theadapter 50000 is a generally planar tab that is coextensive with thetubule 1200. One skilled in the art will appreciate that the adapter50000 need not be coextensive with the tubule and may be constructed ofvarying sizes and shapes depending upon the application. Moreover, morethan one adapter may be provided.

The adapter 50000 may be constructed of any material suitable for use inconstruction the tubule 1200. For example, the adapter may beconstructed of polyurethane. The adapter 50000 may be constructed of thesame or a different material than the tubule 1200. To facilitatehandling, the adapter 50000 may be constructed of a material havingincreased rigidity compared to the material of the tubule 1200, forexample a high durometer polyurethane. In certain embodiments, theadapter 50000 may be manufactured with the tubule 1200 in, for example,a co-extrusion process or an injection molding process. Alternatively,the adapter 50000 may be manufactured independently and attached to thetubule 1200 in a post-forming process by, for example, bonding.

The exemplary embodiment of FIGS. 35A-35E also includes a container 2000and an interface 3000, as described above. The container 2000 removablyand replaceably encloses the tubule 1200 to protect the sample tubule1200 and when removed, may allow direct manipulation of tubule 1200. Aportion of adapter 50000 may not be enclosed by container 2000. Theexposed portion of the adapter 50000 can be directly accessed by a userfor labeling, handling and other processing. The interface 3000 includesan interface channel 3700 that communicates with an opening in thetubule 1200 to facilitate delivery of a sample to the tubule 1200. Inthe illustrated embodiment, a removable and replaceable cover 58600 isprovided to selectively open and close the interface channel 3700. Theexemplary cover 58600 includes a sample collection instrument in theform of a tissue swab 584 for collecting tissue samples from a samplesource.

FIGS. 35A-B illustrate an embodiment of the adapter 50000 that isconstructed to facilitate handling of the sample vessel 1000.

FIG. 35C illustrates an embodiment of the adapter 50000C that isdesigned to facilitate delivery of a reagent or a sample from anexternal device, such as a needle 9000 from a reagent injectorcartridge. The adapter 50000C includes a reversible or irreversiblepressure gate 4800 that provides a fluid channel to permit selectivedisplacement of a fluid, e.g., a sample or reagent, between the tubule1200 and the external device, in the present embodiment, needle 9000.The adapter 50000C may include a self-sealing membrane 54000, valve, orother sealing mechanism to facilitate selective communication with theexternal device. A reservoir 50200 may be provide to contain a fluiddelivered from the external device or fluid from the tubule 1200. Inuse, the needle 9000 may penetrate the self-sealing membrane 54000 todeliver fluid to the reservoir 50200 or to withdraw fluid from thereservoir 50200. Pressure gate 4800 may be opened in the mannerdescribed above, e.g. by compressing the tubule 1200 or the reservoir50200, to withdraw fluid from the reservoir 50200 or to deliver fluid tothe reservoir 50200 from the tubule 1200. The needle 9000 may be coupledwith a sensor, such as an electrode, a fiber optical sensor, forpenetrating the self sealing membrane 54000 and measuring a sampleproperty.

FIG. 35D illustrates an embodiment of the adapter 50000D that comprisesa compressible reservoir 50600 and a reversible or irreversible pressuregate 4800 that provides a fluid channel to permit selective displacementof a fluid, e.g., a sample or reagent, to the tubule 1200 from thecompressible reservoir 50600. The compressible reservoir 50600 maycontain a pre-packed reagent. In certain embodiments, the compressiblereservoir 50600 may be a blister pack. Upon compression of thecompressible reservoir 50600, pressure gate 4800 may open and fluid withthe compressible reservoir 50600 can be displaced in to the tubule 1200.

FIG. 35E illustrates an embodiment of the adapter 50000E that comprisesa reservoir 50200, a first reversible or irreversible pressure gate4800A that provides a fluid channel to permit selective displacement ofa fluid, e.g., a sample or reagent, between the tubule 1200 and thereservoir 50200, and a second reversible or irreversible pressure gate4800B that provides a fluid channel to permit selective displacement ofa fluid, e.g., a sample or reagent, between an external device 50800 andthe reservoir 50200. A connector 50900 may be provided to interface withthe external device 50800. Such device may be an Access™ card forMicronics Inc., a LabChip® product from Caliper, Inc. or a GeneChip®from Affymetrix, Inc.

FIGS. 36A-E illustrate another exemplary embodiment of a sample vessel60000 that comprises a tubule 1000 and an apparatus 60200 for drawing asample into the tubule 1200 of the sample vessel 1000. The apparatus60200 includes a cylindrical housing 60400 having an opening 60600 forreceiving the tubule 1200. The opening 60200 extends from a proximal end60800 to a distal end 61000 of the housing 60400. Both the housing 60400and the opening 60600 can be sized and shaped to accommodate the sizeand shape of the tubule 1200 or other sample vessels. For example, thehousing 60400 and opening 60600 are cylindrical in shape and have acircular cross-section analogous to that of the tubule 1200. The adapter60000 comprises first means 61200 for compressing a first portion of thetubule 1200 and second means 61400 for compressing a second portion ofthe tubule 1200. The first compression means 61200 may be spaced apartfrom the second compression means 61400. For example, the firstcompression means 61200 may be positioned at the proximal end 60800 ofthe housing 60200 and the second compression means 61400 may bepositioned at the distal end 61000 of the housing 60200. The spacingbetween the first and second compression means may be selected based onthe desired sample collection volume in the tubule 1200.

The first compression means 61200 may comprise a first pair of spacedapart rollers, 61600A and 61600B. At least one of the rollers 61600A-Bmay be selectively movable into contact with the other roller tocompress the tubule 1200 between the rollers 61600A-B. A first activator62000 may be coupled to the rollers 61600A, 61600B to effect separationor compression of the rollers. The second compression means 61200 maycomprise a second pair of spaced apart rollers, 61800A and 61800B. Atleast one of the rollers 61800A-B may be selectively movable intocontact with the other roller to compress the tubule 1200 between therollers 61800A-B. A second activator 62200 may be coupled to the rollers61800A, 61800B to effect separation or compression of the rollers. Inaddition to rollers, or other compression mechanisms may be employed forthe first and second compression means, including the compressionmembers described above. Any structure suitable for selectivecompression of the tubule 1200 may be employed. The first and secondcompression means need not be the same structure.

In use, the tubule 1200 is inserted into the opening 60600 at theproximal end 60800 of the housing 60400 and drawn completely through theopening 60600 to the distal end 61000 of the housing 60400. As thetubule 1200 is drawn through the housing 60400, the tubule 1200 isflatten and compressed, as illustrated in FIG. 36B, to evacuate thetubule 1200. At the time of sample collection, a cover 68600 may beremoved to expose a sample collection instrument, such as a needle68400, that is in fluid connection with the tubule 1200. The needle68400 can be inserted into the sample source and the first compressionmeans 61200 may be separated to draw the sample into the tubule 1200, asillustrated in FIG. 36C. The sample vessel 1000 may then be insertedinto a device 63000 for removing the needle 68400, or other samplecollection instrument, as illustrated in FIG. 36D. The device 63000 mayalso include a mechanism for sealing the proximal end of the tubule 1200after the needle 68400 is removed, by, for example, compressing andheating the wall of the tubule 1200 at the proximal end to bond or fusethe walls together. The second compression means 61400 may separate andthe adapter 60000 may be removed from the tubule 1200, as illustrated inFIG. 36E.

While the sample vessels disclosed herein have been particularly shownand described with references to exemplary embodiments thereof, it willbe understood by those skilled in the art that various changes in formand details may be made therein without departing from the spirit andscope of the disclosure. Those skilled in the art will recognize or beable to ascertain using no more than routine experimentation, manyequivalents to the exemplary embodiments described specifically herein.Such equivalents are intended to be encompassed in the scope of thepresent disclosure.

1. A flexible vessel for thermal cycling a sample, the flexible vesselcomprising: a soft-sided reaction vessel comprising: a receptacle forreceiving the sample, and a flexible body coupled to the receptacle, theflexible body having first and second portions coupled together and influid communication with each other, wherein the flexible body isconfigured for insertion into a thermal cycler such that the firstportion is received into a first temperature zone and the second portionis received into a second temperature zone of the thermal cycler.
 2. Thevessel of claim 1, wherein the flexible body is formed from two faces ofplastic sheeting that are sealed together to form sealed sides.
 3. Thevessel of claim 2, further comprising a plurality of additionalsoft-sided reaction vessels, each having a receptacle and a flexiblebody coupled to the receptacle, wherein the plurality of additionalbodies are formed from the two faces of plastic sheeting and each hassealed sides.
 4. The vessel of claim 3, wherein the two faces of plasticsheeting are sealed together such that each flexible body is separatedfrom its adjacent flexible body by a sealed side.
 5. The vessel of claim1, wherein the flexible body contains dried PCR reagents.
 6. A flexiblevessel for thermal cycling a sample, the flexible vessel comprising twofaces of flexible material that are sealed together to form a pluralityof flexible bodies, each flexible body separated from its adjacentflexible body by a sealed side, and each flexible body having a firstportion and a second portion, the first and second portions coupledtogether and in fluid communication with each other, wherein theflexible bodies are configured for insertion into a thermal cyclingapparatus such that the first portion of each flexible body is receivedinto a first temperature zone and the second portion of each flexiblebody is received into a second temperature zone of the thermal cycler.7. The vessel of claim 6, further comprising a plurality of receptacles,each receptacle fluidly coupled to its respective flexible body.
 8. Thevessel of claim 6, wherein the flexible material is a thin plastic film.9. The vessel of claim 8, wherein the thin plastic film comprisesaluminum lamination.
 10. The vessel of claim 8, wherein the thin plasticfilm is selected from the group consisting of polyethylene, polyurethaneand alloys thereof.
 11. The vessel of claim 8, wherein the thin plasticfilm transmits about 80% to about 90% of incident light.
 12. The vesselof claim 6, wherein the vessel body has a coefficient of heat transferof about 0.02 to about 20 W/m*degK.
 13. The vessel of claim 6, whereineach flexible body contains dried PCR reagents.
 14. A thermal cyclingsystem comprising the reaction vessel of claim 6 received into thethermal cycler, wherein the first temperature zone of the thermal cycleris configured for receiving the first portion of the reaction vessel,the first temperature zone comprising a first heater, the firsttemperature zone movable between an open orientation in which the firstheater affects the temperature of the reaction mixture contained withinthe first portion, and a closed orientation in which the reactionmixture is forced from the first portion and the first heater does notsubstantially affect the temperature of the reaction mixture, and thesecond temperature zone of the thermal cycler is configured forreceiving the second portion of the reaction vessel, the secondtemperature zone comprising a second heater, the second temperature zonemovable between an open orientation in which the second heater affectsthe temperature of the reaction mixture contained within the secondportion, and a closed orientation in which the reaction mixture isforced from the second portion and the second heater does notsubstantially affect the temperature of reaction mixture.
 15. Thethermal cycling system of claim 14, wherein each flexible body containsa PCR reaction mixture.
 16. The thermal cycling system of claim 15,wherein the PCR reaction mixture is sealed within each flexible body.17. The thermal cycling system of claim 16, wherein each flexible bodyis heat-sealed to seal the reaction mixture within each flexible body.18. The thermal cycling system of claim 16, wherein the first portion ofeach flexible body is coupled to a fitting, and closure of the fittingseals the reaction mixture within its respective flexible body.
 19. Thevessel of claim 1, wherein the flexible body comprises a thin plasticfilm.
 20. The vessel of claim 19, wherein the flexible body furthercomprises aluminum lamination.
 21. The vessel of claim 19, wherein thethin plastic film is selected from the group consisting of polyethylene,polyolefin, polyurethane and alloys thereof.
 22. The vessel of claim 19,wherein the thin plastic film transmits about 80% to about 90% ofincident light.
 23. The vessel of claim 1, wherein the vessel body has acoefficient of heat transfer of about 0.02 to about 20 W/m*degK.
 24. Thevessel of claim 1, wherein the flexible body is formed from two walls ofplastic and has a flattenable cross-sectional profile.
 25. The vessel ofclaim 24, further comprising a plurality of additional soft-sidedreaction vessels, each having a receptacle and a flexible body coupledto the receptacle, wherein the plurality of additional bodies are eachformed from two walls of plastic and has a flattenable cross-sectionalprofile.
 26. A flexible vessel for thermal cycling a sample, theflexible vessel comprising a plurality of flexible bodies, each flexiblebody separated from its adjacent flexible body, and each flexible bodyhaving a first portion and a second portion, the first and secondportions coupled together and in fluid communication with each other,wherein the flexible bodies are configured for insertion into a thermalcycling apparatus such that the first portion of each flexible body isreceived into a first temperature zone and the second portion of eachflexible body is received into a second temperature zone of the thermalcycler.
 27. A thermal cycling system comprising the reaction vessel ofclaim 26 received into the thermal cycler, wherein a first temperaturezone of the thermal cycler is configured for receiving the firstportions of the flexible bodies of the reaction vessel, the firsttemperature zone comprising a first heater, the first temperature zonemovable between an open orientation in which the first heater affectsthe temperature of reaction mixtures contained within the respectivefirst portions, and a closed orientation in which the reaction mixturesare forced from the respective first portions and the first heater doesnot substantially affect temperatures of the reaction mixtures, and asecond temperature zone of the thermal cycler is configured forreceiving the second portions of the flexible bodies of the reactionvessel, the second temperature zone comprising a second heater, thesecond temperature zone movable between an open orientation in which thesecond heater affects temperatures of the reaction mixtures containedwithin the respective second portion, and a closed orientation in whichthe reaction mixtures are forced from the respective second portions andthe second heater does not substantially affect temperatures of thereaction mixtures.
 28. The thermal cycling system of claim 27, whereineach flexible body is sealed to seal the reaction mixture within eachflexible body.